Muscarinic agonists

ABSTRACT

The present invention relates to compounds of Formula (I): which are agonists of the M-1 muscarinic receptor

This is the national phase application, under 35 U.S.C. §371, forPCT/US2004/005234 filed on 12 Mar. 2004, which claims the priority ofU.S. Provisional Application No. 60/456,911 filed on 21 Mar. 2003.

The present invention relates to the field of pharmaceutical and organicchemistry and provides compounds that are active at the muscarinicreceptors.

The compounds of the present invention are muscarinic agonists. Morespecifically, the compounds of the present invention are selectiveagonists of the muscarinic M-1 receptor. As such, they are useful fortreating a variety of disorders of the central nervous system and otherbody systems. These disorders include cognitive disorders, ADHD,obesity, Alzheimer's disease, psychoses including schizophrenia, and foralleviation of intraocular pressure such as that found in glaucoma.

Certain indane-like compounds are described as useful for treatingconditions associated with malfunctioning of the muscarinic cholinergicsystem in PCT Publication Nos. WO 97/25983, published 24 Jul. 1997, andWO 99/04778, published 4 Feb. 1999.

The present invention provides compounds of Formula I:

wherein

-   -   Q, X, Y, and Z are independently selected from the group        consisting of CR¹ and N, provided that no more than two of Q, X,        Y, and Z are N and at least two of Q, X, Y, and Z are CH; or Y        is CH, Z is CH, and the moiety “Q═X” represents “S” to form a        thiophene ring;    -   R¹ is independently at each occurrence selected from the group        consisting of hydrogen, halogen, C₁-C₄ alkoxy, and C₁-C₄ alkyl;    -   R² is selected from the group consisting of halogen; C₁-C₄        alkoxy; C₁-C₄ alkyl; C₃-C₈ cycloalkyl; cyano; trifluoromethyl;        pyridinyl optionally substituted with one to two substituents        independently selected from the group consisting of halogen,        C₁-C₄ alkoxy, and C₁-C₄ alkyl; thienyl optionally substituted        with one substituent selected from the group consisting of        halogen, C₁-C₄ alkoxy, and C₁-C₄ alkyl; phenyl optionally        substituted with from one to three substituents independently        selected from the group consisting of halogen, C₁-C₄ alkoxy,        C₁-C₄ alkyl, trifluoromethyl, and cyano; and pyrrolyl optionally        substituted with one to two substituents independently selected        from the group consisting of halogen, C₁-C₄ alkoxy, and C₁-C₄        alkyl;    -   R³ is a radical of the formula        (Z)-(Y)—(X)—        -   wherein        -   X is selected from the group consisting of

and a straight-chain C₁-C₄ alkandiyl optionally substituted with methyl,geminal dimethyl, or phenyl;

-   -   -   Y is selected from the group consisting of O and S; and        -   Z is selected from the group consisting of C₁-C₆ alkyl;            C₃-C₈ cycloalkyl optionally substituted with one to three            substituents independently selected from the group            consisting of halogen, C₁-C₄ alkoxy, C₁-C₄ alkyl,            trifluoromethyl, cyano, and nitro; phenyl optionally            substituted with one to three substituents independently            selected from the group consisting of halogen, C₁-C₄ alkoxy,            C₁-C₄ alkyl, trifluoromethyl, cyano, and nitro; naphthyl            optionally substituted with one to three substituents            independently selected from the group consisting of halogen,            C₁-C₄ alkoxy, C₁-C₄ alkyl, trifluoromethyl, cyano, and            nitro; heteroaryl optionally substituted with one or two            substituents independently selected from the group            consisting of halogen, C₁-C₄ alkoxy, and C₁-C₄ alkyl; and            heterocycle optionally substituted with one or two            substituents independently selected from the group            consisting of halogen, C₁-C₄ alkoxy, and C₁-C₄ alkyl;

    -   R^(a) is selected from the group consisting of hydrogen and        methyl;

    -   or R³ and R^(a) are taken together with the nitrogen with which        they are attached to form a heterocycle optionally substituted        with one or two substituents independently selected from the        group consisting of halogen, C₁-C₄ alkoxy, and C₁-C₄ alkyl;

    -   R⁴ is selected from the group consisting of hydrogen, hydroxy,        and fluoro;

    -   R⁵ is selected from the group consisting of hydrogen, halogen,        C₁-C₄ alkoxy, and C₁-C₄ alkyl;

    -   R^(b) is selected from the group consisting of hydrogen, methyl,        and ethyl; and

    -   m is one or two;

    -   or pharmaceutically acceptable addition salts thereof.

The present invention also provides pharmaceutical compositions,comprising a compound of Formula I and a pharmaceutically acceptablediluent.

Because the compounds of Formula I are agonists of the M-1 muscarinicreceptor, the compounds of Formula I are useful for the treatment of avariety of disorders associated with muscarinic receptors, including:cognitive disorders (including age-related cognitive disorder, mildcognitive impairment, cognitive impairment associated withschizophrenia, and chemotherapy-induced cognitive impairment), ADHD,mood disorders (including depression, mania, bipolar disorders),psychosis (in particular schizophrenia), dementia (including Alzheimer'sdisease, AIDS-induced dementia, vascular dementia, and dementia lackingdistinctive histology), Parkinson's disease, and Huntington's Chorea.Also, the present compounds are useful for treating chronic colitis,including Crohn's disease. Additionally, the present compounds areuseful for the treatment of pain (including acute pain and chronicpain), xerostomia (dry mouth), Lewy body disease (including diffuse Lewybody disease), aphasia (including primary aphasia and primary aphasiasyndromes), and hypotensive syndromes.

In another embodiment the present invention provides methods of treatingdisorders associated with muscarinic receptors, comprising:administering to a patient in need thereof an effective amount of acompound of Formula I. That is, the present invention provides for theuse of a compound of Formula I or a pharmaceutical composition thereoffor the manufacture of a medicament for the treatment of disordersassociated with muscarinic receptors. The present invention alsoprovides a compound of Formula I for use in therapy.

As used herein, the following terms have the meanings indicated:

The term “halo” or “halogen” refers to a chloro, fluoro, bromo or iodoatom.

The term “C₁-C₄ alkyl” refers to a straight or branched alkyl chainhaving from one to four carbon atoms, examples of which include methyl,ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, and t-butyl.The term “C₁-C₄ alkandiyl” refers to a straight- or branched-chainalkandiyl having from one to four carbon atoms in total, examples ofwhich include methylene, ethylene, tetramethylene,1-methylpropan-1,3-diyl, 2-methylpropan-1,3-diyl, and butan-2,3-diyl.The term “C₃-C₈ cycloalkyl” refers to cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

The term “C₁-C₄ alkoxy” refers to a straight or branched alkyl chainhaving from one to four carbon atoms attached to an oxygen atom,examples of which include methoxy, ethoxy, n-propoxy, iso-propoxy,n-butoxy, iso-butoxy, sec-butoxy, and t-butoxy.

The term “heteroaryl” is taken to mean a stable unsaturated five- orsix-membered ring containing from 1 to 2 heteroatoms selected from thegroup consisting of nitrogen, oxygen and sulfur. Examples of heteroarylinclude pyridinyl, pyrimidinyl, pyrazinyl, pyrrolyl, oxazolyl,isoxazolyl, imidazolyl, thiazolyl, pyridazinyl, furyl, thienyl, and thelike. Preferred heteroaryl groups are thienyl, pyridinyl, and furyl.

The term “heterocycle” is taken to mean a stable saturated five- orsix-membered ring containing from 1 to 3 heteroatoms selected from thegroup consisting of nitrogen, oxygen and sulfur. Examples of heterocycleinclude pyrrolidinyl, piperidinyl, piperazinyl, tetrahydrofuryl,morpholino, and the like

The compounds of the present invention form pharmaceutically acceptableacid addition salts with a wide variety of organic and inorganic acidsand include the physiologically acceptable salts which are often used inpharmaceutical chemistry. Such salts are also part of this invention. A“pharmaceutically-acceptable addition salt” is formed from apharmaceutically-acceptable acid as is well known in the art. Such saltsinclude the pharmaceutically acceptable salts listed in Journal ofPharmaceutical Science, 66, 2-19 (1977) which are known to the skilledartisan. Typical inorganic acids used to form such salts includehydrochloric, hydrobromic, hydriodic, nitric, sulfuric, phosphoric,hypophosphoric, metaphosphoric, pyrophosphoric, and the like. Saltsderived from organic acids, such as aliphatic mono and dicarboxylicacids, phenyl substituted alkanoic acids, hydroxyalkanoic andhydroxyalkandioic acids, aromatic acids, aliphatic and aromatic sulfonicacids, may also be used. Such pharmaceutically acceptable salts thusinclude chloride, bromide, iodide, nitrate, acetate, phenylacetate,trifluoroacetate, acrylate, ascorbate, benzoate, chlorobenzoate,dinitrobenzoate, hydroxybenzoate, methoxybenzoate, methylbenzoate,o-acetoxybenzoate, isobutyrate, phenylbutyrate, α-hydroxybutyrate,butyne-1,4-dicarboxylate, hexyne-1,4-dicarboxylate, caprate, caprylate,cinnamate, citrate, formate, fumarate, glycollate, heptanoate,hippurate, lactate, malate, maleate, hydroxymaleate, malonate,mandelate, mesylate, nicotinate, isonicotinate, oxalate, phthalate,teraphthalate, propiolate, propionate, phenylpropionate, salicylate,sebacate, succinate, suberate, benzenesulfonate,p-bromobenzenesulfonate, chlorobenzenesulfonate, ethylsulfonate,2-hydroxyethylsulfonate, methylsulfonate, naphthalene-1-sulfonate,naphthalene-2-sulfonate, naphthalene-1,5-sulfonate, p-toluenesulfonate,xylenesulfonate, tartrate, and the like.

The present invention includes the stereoisomers and tautomers of thecompounds of Formula I. Herein, the Cahn-Prelog-Ingold designations of(R)— and (S)— and the cis and trans designation of relativestereochemistry are used to refer to specific isomers and relativestereochemistry.

As with any group of pharmaceutically active compounds, some groups arepreferred in their end use application. The following paragraphs definepreferred classes.

-   -   a) When R⁴ is not hydrogen, compounds which have trans        stereochemistry at the 1- and 2-position are preferred.    -   b) When R⁴ is not hydrogen, compounds which have the trans        stereochemistry at the 1- and 2-position shown below are more        preferred.

-   -   c) R^(a) is methyl.    -   d) R⁵ is hydrogen.    -   e) R⁴ is hydroxy.    -   f) m is one.    -   g) R^(a) is methyl, R⁵ is hydrogen, R⁴ is hydroxy, and m is one.    -   h) Q, X, Y, and Z are each CR¹ provided that at least two of Q,        X, Y, and Z are CH.    -   i) R¹ is hydrogen.    -   j) R¹ is halogen.    -   k) R¹ is fluoro.    -   l) Q, X, Y, and Z are each CH.    -   m) One of Q, X, Y, and Z is CF and the others are CH.    -   n) Q is CF and X, Y, and Z are each CH.    -   p) R² is phenyl optionally substituted with from one to three        substituents independently selected from the group consisting of        halogen, C₁-C₄ alkoxy, C₁-C₄ alkyl, trifluoromethyl, and cyano.    -   q) R² is phenyl.    -   r) X is a straight-chain C₁-C₄ alkandiyl.    -   s) Y is O.    -   t) Y is S.    -   u) Z is C₁-C₄ alkyl.    -   v) Z is phenyl optionally substituted with one to three        substituents independently selected from the group consisting of        halogen, C₁-C₄ alkoxy, C₁-C₄ alkyl, trifluoromethyl, cyano, and        nitro.        The preceding paragraphs may be combined to define additional        preferred classes of compounds.

The compounds of Formula I in which R⁴ is hydroxy are prepared byprocedures described in Scheme A. In Scheme A all substituents, unlessotherwise indicated, are as previously defined, and all reagents arewell known and appreciated in the art.

In Scheme A, step a, the compound of Formula (1) is resolved to give asubstantially pure compound of Formula (2). The compound of Formula (1)is readily prepared by methods well known and appreciated in the art,such as those found in PCT Publication Nos. WO 97/25983, published 24Jul. 1997; and WO 99/04778, published 4 Feb. 1999. As used herein theterm “substantially pure” refers to enantiomeric purity. The desiredstereochemistry in final compounds of Formula I may be convenientlyintroduced in Scheme A, step a, by resolution of compounds of Formula(1). Further processing of resolved compounds of Formula (1), via stepsb, c, d, and optional step e, described infra, will result insubstantially pure compounds of Formula I. Substantially pure compoundsof Formula I can be prepared which are greater than 80%, preferablygreater than 90%, more preferably greater than 95%, most preferablygreater than 97% enantiomerically pure. The compound of Formula (1) canbe resolved by chiral chromatography or by fractional crystallization ofdiasteriomeric acid addition salts. It is expected that a wide varietyof such salts are suitable for this purpose. In practice, isomers ofmandelic acid have been found to be particularly useful.

For example, the compound of Formula (1) is contacted with the selectedacid. Generally, from about 0.4 molar equivalents to a large excess ofthe selected acid can be used with about 0.4 to 1.5 molar equivalentsbeing preferred and with about 0.5 to 1.1 molar equivalents being morepreferred. The resolution is typically carried out by crystallizing theacid addition salt from a solution. In particular, solvents such aslower alcohols, including methanol are useful. It may be advantageous touse small amounts of water with the selected solvent(s) in order tocarry out the resolution in a reasonable volume. The use of ananti-solvent may also be advantageous. As used herein, the term“anti-solvent” refers to a solvent in which the salt is significantlyless soluble compared to the other selected solvent(s). Preferably, whenan anti-solvent is used it is miscible with the other selectedsolvent(s). Suitable anti-solvents include ethers, such as diethylether, methyl t-butyl ether, and the like, and lower alkyl acetates,such as methyl acetate, ethyl acetate, isopropyl acetate, propylacetate, iso-butyl acetate, sec-butyl acetate, butyl acetate, amylacetate, iso-amyl acetate, and the like, and alkanes, such as pentane,hexane, heptane, cyclohexane, and the like. When the racemic mixture isused, care should be taken in using an anti-solvent to avoidcrystallization of the salt of the undesired diastereomeric salt.

Typically, the crystallization is carried out at initial temperatures ofabout 40° C. to reflux temperature of the selected solvent(s). Themixture is then cooled to give the salt. Seeding may be advantageous.Preferably the crystallization solution is cooled slowly. Thecrystallization is most conveniently cooled to temperatures of ambienttemperature to about −20° C. The salt can be collected using techniquesthat are well known in the art, including filtration, decanting,centrifuging, evaporation, drying, and the like. The compound of Formula(2) can be used directly as the acid addition salt of the selected acid.Alternately, before use the compound of Formula (2) can be isolated asanother acid addition salt after acid exchange or can by isolated as thebase by extraction under basic conditions as is well known andappreciated in the art.

As is readily apparent to one skilled in the art the depicted compoundof Formula (2) is of the trans configuration at the 1- and 2-positionsof the indane nucleus. Cis compounds are readily prepared from suchtrans compounds by protection of the amine, inversion of the hydroxycenter, followed by deprotection as needed. There are numerous methodswhich allow for inversions of hydroxy centers, such as by Mitsunobureaction with suitable carboxylic acids, including acetic acid andbenzoic acid, followed by hydrolysis. Alternately, an appropriatelyresolved amino-indanol may be selectively nitrated to produce a compoundof Formula (2). For example, the resolved amino-indanol may beintroduced to a nitrating agent, such as nitric acid or sodium nitrate.This reaction may be conducted in the presence of a strong acid, such astrifluoroacetic acid or sulfuric acid. Subsequently, the reaction may beneutralized with an appropriate base such as sodium hydroxide. Methodsof nitration are well known in the art; see, for example, OrganicChemistry, Morrison & Boyd, 5th Ed (Allyn & Bacon, Inc.).

Reaction Scheme A, step b, depicts the formation of a compound ofFormula (3). It is understood that the compound of Formula (3) can beone in which R is a group as desired in the final product of Formula Ias defined above. R may also combine with the carbonyl to form aprotecting group, such as t-BOC, which can be later removed beforeincorporation of an R group as desired in the final product of FormulaI. The selection and use of suitable protecting groups is well known andappreciated in the art (Protecting Groups in Organic Synthesis, TheodoraGreene (Wiley-Interscience)).

For example, where R is a group as desired in the final product, thecoupling reaction depicted in step b is carried out using theappropriate acid or the acid halide derived therefrom. Appropriate acidsinclude various substituted benzoic acids and acid halides, heteroarylacids and acid halides, and various biaryl carboxylic acids and acidhalides. Examples include biphenyl carboxylic acid and3-fluorobiphenyl-4-carboxylic acid.

For example, the compound of Formula (2) is contacted with anappropriate acid to give a compound of Formula (3). Such couplingreactions are common in peptide synthesis and synthetic methods usedtherein can be employed. For example, well known coupling reagents, suchas resin-bound reagents and carbodiimides with or without the use ofwell-known additives such as N-hydroxysuccinimide,1-hydroxybenzotriazole, etc. can be used to facilitate this acylation.The reaction is conventionally conducted in an inert aprotic polardiluent such as dimethylformamide (DMF), methylene chloride(dichloromethane), chloroform, acetonitrile, tetrahydrofuran (THF), andthe like. Typically the reaction is carried out at temperatures of fromabout 0° C. to about 60° C. and typically require from about 1 to about24 hours. Upon reaction completion, the product of Formula (3) isrecovered by conventional methods including extraction, precipitation,chromatography, filtration, trituration, crystallization and the like.

Alternatively, for example, the compound of Formula (2) is contactedwith an acid halide of an appropriate acid to give a compound of Formula(3). Such acid halides are commercially available or readily preparedfrom the corresponding acids by methods well known in the art, such asby the action of phosphorous trichloride, phosphorous tribromide,phosphorous oxychloride, phosphorous pentachloride, thionyl chloride,thionyl bromide, or oxalyl chloride, with or without a small amount ofdimethylformamide, in an inert solvent such as, toluene, methylenechloride, or chloroform; at temperatures of from about 0-80° C. Thereaction is typically carried out for a period of time ranging from 1hour to 24 hours. The acid halide can be isolated and purified or canoften be used directly, that is, with or without isolation and/orpurification. The coupling reactions generally use a suitable base toscavenge the acid generated during the reaction. Suitable bases include,by way of example, sodium hydroxide, potassium hydroxide, pyridine,triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, and thelike. The reaction is conventionally conducted in a solvent such asmethylene chloride, chloroform, tetrahydrofuran and the like, or underSchotten-Baumann conditions in a solvent mixture such as methylenechloride, ethyl acetate, toluene and water. Typically the couplingreaction is carried out at temperatures of from about −20° C. to about80° C. and typically require from about 1 to about 24 hours. Uponreaction completion, the product of Formula (3) is recovered byconventional methods including extraction, precipitation,chromatography, filtration, trituration, crystallization and the like.

Reaction Scheme A, step c, depicts the reduction of a nitro group togive a compound of Formula (4). Such reductions can be carried out by avariety of methods that are well known in the art.

For example, a compound of Formula (3) may be hydrogenated over acatalyst, such as palladium-on-carbon, to give a compound of Formula(4). Such hydrogenations are generally carried out in a solvent and avariety of solvents are suitable, for example methanol, ethanol,isopropanol, tetrahydrofuran, or ethyl acetate or mixtures thereof. Thehydrogenation may be performed at an initial hydrogen pressure of 20-180psi (137-1241 kPa). The reaction is typically carried out at temperatureof about 0° C. to about 60° C. The reaction typically requires 1 hour to3 days. The product can be isolated and purified by techniques wellknown in the art, such as filtration, extraction, evaporation,trituration, precipitation, chromatography, and recrystallization.

In Scheme A, step d, a compound of Formula (4) is contacted with anappropriate amidine-forming agent to give a compound of Formula I.Appropriate amidine-forming agents include1-methylthio-1-methyl-N-(4-fluorobenzyl)-N-methylimmonium triflate and1-methylthio-1-methyl-N-(4-fluorobenzyl)-N-methylimmonium iodide. One ofordinary skill in the art will recognize that appropriateamidine-forming agents may be prepared in advance or in situ if desired.

For example, a compound of Formula (4) is contacted with from about 1-3equivalents of an appropriate amidine-forming agent. The reaction isgenerally carried out in a dry solvent such as methylene chloride,toluene, or tetrahydrofuran at temperatures of from about −20° C. to 50°C. The reaction is carried out using an appropriate base such aspyridine, collidine, or triethylamine. The reaction typically requires 1to 18 hours. The product can be isolated and purified by techniques wellknown in the art, such as quenching, filtration, extraction,evaporation, trituration, precipitation, chromatography, andrecrystallization.

As will be readily appreciated, where R is a protecting group introducedin step b, the protecting group can be removed after step d and theresulting amine coupled with an appropriate acid or acid halide as alsodescribed above in step b to give a compound of Formula I.

Some compounds of Formula I are intermediates for other final compoundsof Formula I. For example, when R² is iodo, another reagent, forexample, 2-(tributylstannyl)thiophene or 2-(tributylstannyl)pyridine,may be used to displace iodo as a leaving group and substitute adifferent R² group as desired in the final product.

In Scheme A, optional step e, not shown, an acid addition salt of acompound of Formula I is formed using a pharmaceutically-acceptableacid. The formation of acid addition salts is well known and appreciatedin the art.

The compounds of Formula I in which R⁴ is hydrogen are prepared fromcompounds of Formula (3) or from amine protected compounds of Formula(2) by deoxygenation. Such deoxygenation reactions are readily carriedout using procedures well known in the art, described, for example, byLarock, Comprehensive Organic Transformations, pg. 44-52 (1999).Alternately, the compounds of Formula I in which R⁴ is hydrogen areprepared by procedures described in Scheme B. In Scheme B allsubstituents, unless otherwise indicated, are as previously defined, andall reagents are well known and appreciated in the art.

Reaction Scheme B, step a, depicts the reductive amination of a compoundof Formula (5) to give a compound of Formula (6). Such reductiveaminations are carried out under a variety of conditions. The reactiondepicted in Scheme B, step a, can be carried out using ammonia or aprotected amine, such as benzyl amine, dibenzyl amine, and the likefollowed by deprotection to give the compound of Formula (6).

For example, a compound of Formula (5) is reacted with an excess ofammonia and sodium cyanoborohydride to give a compound of Formula (6).As is well known in the art, it may be advantageous to monitor andadjust the pH during such reactions. The reaction is carried out in asolvent, such as methanol, ethanol, isopropanol, and water or mixturesthereof. Typically the reaction is carried out at temperatures of fromabout 0° C. to about 60° C. and typically require from about 1 to about24 hours. Upon reaction completion, the product of Formula (6) isrecovered by conventional methods including extraction, precipitation,chromatography, filtration, trituration, crystallization, and the like.

Reaction Scheme B, steps b, c, d, and optional step e, are carried outby the methods described in Scheme A, steps b, c, d, and optional stepe, to give a compound of Formula I.

The compounds of Formula I in which R⁴ is fluoro are prepared fromcompounds of Formula (3) or from amine protected compounds of Formula(2) by halogenation procedures well known in the art, described, forexample, by Larock, Comprehensive Organic Transformations, pg. 689-701(1999).

The present invention is further illustrated by the following examplesand preparations. These examples and preparations are illustrative onlyand are not intended to limit the invention in any way.

The terms used in the examples and preparations have their normalmeanings unless otherwise designated. For example, “° C.” refers todegrees Celsius; “M” refers to molar or molarity; “mmol” refers tomillimole or millimoles; “g” refers to gram or grams; “mL” refersmilliliter or milliliters; “mp” refers to melting point; “brine” refersto a saturated aqueous sodium chloride solution; etc. In the ¹H NMR, allchemical shifts are given in δ, unless otherwise indicated.

Coupling Procedures

Method A 2′-Chlorobiphenyl-4-carboxylic acid

Combine methyl-4-bromobenzoate (1.0 g, 4.65 mmol), 2-chlorophenylboronicacid (799 mg, 5.1 mmol), Pd(OAc)₂ (51 mg, 0.46 mmol) and sodiumcarbonate (1.5 g, 13.9 mmol) in DMF (20 mL) and water (2.0 mL) withstirring. Purge the reaction mixture with argon, add triphenylphosphine(61 mg, 0.23 mmol) and purge again with argon. Place the sealed reactionin an oil bath maintained at 80° C. and allow to stir for 1 hour. Coolthe reaction to room temperature, dilute with ethyl acetate and filterthrough a short plug of celite with additional ethyl acetate. Wash theorganics with water, dry over MgSO₄, filter and evaporate. Purificationby flash column chromatography yields 2′-chlorobiphenyl-4-carboxylicacid methyl ester as a yellow solid. Dissolve the purified ester in THF(0.25M) and add an equal volume of 1M NaOH. Stir vigorously at roomtemperature for 15 hours. Upon completion, acidify the reaction withconc. HCl and extract with ethyl acetate. Evaporation of the solventyields 762 mg (67%) of the title compound. MS (ES): m/z 231.1 (M−H).

The following compounds are prepared essentially as described above.

6-(2-Chlorophenyl)pyridine-3-carboxylic MS (ES): m/z 233.9(M + H) acid6-(2,4-Difluorophenyl)pyridine-3- MS (ES): m/z 235.9(M + H) carboxylicacid 6-Phenylpyridine-3-carboxylic acid MS (ES): m/z 214.1(M + H) methylester 6-(2-Methylphenyl)pyridine-3-carboxylic MS (ES): m/z 214.0(M + H)acid 2′-Trifluoromethylbiphenyl-4-carboxylic MS (ES): m/z 265.2(M − H)acid 2-Methylbiphenyl-4-carboxylic acid MS (ES): m/z 211.3(M − H)3-Fluorobiphenyl-4-carboxylic acid MS (ES): m/z 215.1(M − H)2′,6′-Dichlorobiphenyl-4-carboxylic acid MS (ES): m/z 264.9(M − H)2′,6′-Difluorobiphenyl-4-carboxylic acid MS (ES): m/z 233.1(M − H)2′-Methoxybiphenyl-4-carboxylic acid MS (ES): m/z 227.0(M − H)3,4′-Difluorobiphenyl-4-carboxylic acid MS (ES): m/z 233.1(M − H)3,2′-Difluorobiphenyl-4-carboxylic acid MS (ES): m/z 233.1(M − H)3-Chlorobiphenyl-4-carboxylic acid MS (ES): m/z 231.1(M − H)4-(Thien-2-yl)phenyl-1-carboxylic acid MS (ES): m/z 203.1(M − H)4′-Fluorobiphenyl-4-carboxylic acid MS (ES): m/z 214.9(M − H)(Hydrolysis in dioxane at 60° C.) 3′-Fluorobiphenyl-4-carboxylic acid MS(ES): m/z 215.0(M − H) (Hydrolysis in dioxane)3′-Cyanobiphenyl-4-carboxylic acid MS (ES): m/z 222.0(M − H) (Hydrolysiswith LiOH in dioxane)

Method B 5-Phenylpyrazine-2-carboxylic acid

Combine 5-chloropyrazine-2-carboxylic acid methyl ester (626 mg, 3.64mmol), phenylboronic acid (666 mg, 5.45 mmol), cesium fluoride (55 mg,0.36 mmol) and Na₂CO₃ (964 mg, 9.09 mmol) in DMF (5 mL) and water (5 mL)with stirring. Place the hetereogeneous reaction mixture, open to theair, in an oil bath maintained at 80° C. After 5 minutes of heating, addPd(OAc)₂ (81 mg 0.36 mmol) in one portion and stir until reaction turnsblack. Cool the reaction to room temperature, dilute with ethyl acetate,and filter through a short plug of celite with additional ethyl acetate.Wash the organics with water, dry over MgSO₄, filter and evaporate.Purification by flash column chromatography yields2-phenylpyrimidine-5-carboxylic acid methyl ester as a yellow solid.Dissolve the purified ester in THF (0.25M) and add an equal volume of 1MNaOH. Stir vigorously at room temperature for 15 hours. Upon completion,acidify the reaction with conc. HCl and extract with ethyl acetate.Evaporation of the solvent yields 63 mg (8%) of the title compound. ¹HNMR (DMSO): 9.37 (s, 1H), 9.21 (s, 1H), 8.23-8.21 (m, 2H), 7.57-7.77 (m,3H).

The following compounds are prepared essentially as described above.

2′-Fluoro-6′-trifluoromethylbiphenyl-4- MS (ES): m/z 283.1(M − H)carboxylic acid 3,2′,4′-Trifluorobiphenyl-4-carboxylic acid MS (ES): m/z251.1(M − H) 4′-Fluoro-2′-methoxybiphenyl-4-carboxylic MS (ES): m/z245.1(M − H) acid 3-Chloro-2′,4′-difluorobiphenyl-4- MS (ES): m/z267.1(M − H) carboxylic acid 4′-Fluoro-2′-methylbiphenyl-4-carboxylic MS(ES): m/z 229.0(M − H) acid 4′-Trifluoromethylbiphenyl-4-carboxylic MS(ES): m/z 265.1(M − H) acid 2-Fluoro-4-(thien-2-yl)phenyl-1-carboxylicMS (ES): m/z 221.1(M − H) acid

Method C 3′,4′-Difluorobiphenyl-4-carboxylic acid

Combine 3,4-difluorobenzeneboronic acid (1.0 g, 5.2 mmol),methyl-4-bromobenzoate (0.241 g, 1.73 mmol), Pd(OAc)₂ (0.019 g, 0.086mmol), tetrabutylammonium bromide (0.111 g, 0.345 mmol), and potassiumphosphate (0.733 g, 3.454 mmol). Purge the reaction vessel with argonand add anhydrous DMF (20 mL) to the reaction mixture. Heat the sealedreaction vessel to 120° C. with stirring until completion. Cool thereaction to room temperature, dilute with ethyl acetate, and filterthrough a short plug of celite with additional ethyl acetate. Washorganics with water, dry over MgSO₄, filter, and evaporate. Purificationby flash column chromatography yields3′,4′-difluorobiphenyl-4-carboxylic acid methyl ester as a yellow solid.Dissolve the purified ester in dioxane (45 mL) and add an equal volumeof 1 M aqueous NaOH. Heat the reaction vessel to 60° C. with stirringuntil completion. Remove the solvent by evaporation. Dissolve theresidue in dichloromethane and wash with 1N aqueous hydrochloric acid.Dry the organics over MgSO₄, filter and evaporate to yield 0.048 g (12%)of the title compound. MS (ES): m/z 235 (M+H).

The following compounds are prepared essentially as described above.

6-(2-Fluorophenyl)pyridine-3-carboxylic MS (ES): m/z 218.0(M + H) acid3′,5′-Dimethylbiphenyl-4-carboxylic acid MS (ES): m/z 225.0(M − H)3′,5′-Difluorobiphenyl-4-carboxylic acid MS (ES): m/z 233.0(M − H)3′,5′-Dichlorobiphenyl-4-carboxylic acid MS (ES): m/z 267.1(M⁺)3′-Chlorobiphenyl-4-carboxylic acid MS (ES): m/z 230.9(M − H)2′,3′-Difluorobiphenyl-4-carboxylic acid MS (ES): m/z 264.9(M − H)4′-Chlorobiphenyl-4-carboxylic acid MS (ES): m/z 230.9(M − H)

Method D 2′,4′,6′-Trimethylbiphenyl-4-carboxylic acid

Combine 1-iodo-2,4,6-trimethylbenzene (2.966 g, 12.05 mmol),4-carboxyphenylboronic acid (1.0 g, 6.026 mmol), Pd(OAc)₂ (0.0067 g,0.005 mmol), tetrabutylammonium bromide (0.388 g, 1.206 mmol), andpotassium phosphate (2.557 g, 12.05 mmol). Purge the reaction vesselwith argon and add anhydrous DMF (20 mL) to the reaction mixture. Heatthe sealed reaction vessel to 120° C. with stirring until completion asdetermined by TLC. Cool reaction mixture to room temperature. Add methyliodide (1.0 mL, 36.63 mmol) to reaction mixture with continued stirringuntil completion. Dilute the reaction with ethyl acetate and filterthough a short plug of celite with additional ethyl acetate. Wash theorganics with water, dry over MgSO₄, filter and evaporate. Purificationby flash column chromatography yields2′,4′,6′-trimethylbiphenyl-4-carboxylic acid methyl ester as a yellowsolid. Dissolve the purified ester in dioxane (45 mL) and water (5 mL)containing 5 eq of LiOH with stirring at 60° C. Upon completion,evaporate the solvent, acidify the reaction mixture with hydrochloricacid, and extract with ethyl acetate. Dry the organics over MgSO₄,filter, and evaporate to yield 0.023 g (16%) of the title compound. MS(ES): m/z 239.1 (M−H).

The following compounds are prepared essentially as described above.

2′,4′,6′-Trifluorobiphenyl-4-carboxylic acid MS (ES): m/z 251.0(M − H)2′-Fluoro-4′-trifluoromethylbiphenyl-4- MS (ES): m/z 283.0(M − H)carboxylic acid

Method E 2′,4′-Difluorobiphenyl-4-carboxylic acid

Combine 4-carbomethoxyphenylboronic acid (1.021 g, 5.67 mmol),1-bromo-2,4-difluorobenzene (1.000 g, 5.181 mmol.), Pd(OAc)₂ (0.113 g,0.50 mmol), triphenylphosphine (0.149 g, 0.505 mmol), and sodiumcarbonate (1.664 g, 0.568 mmol). Purge the reaction vessel with argon.Add DMF (20 mL) and water (2.0 mL) with stirring. Place sealed reactionin an 80° C. oil bath and allow to stir for 24 hours. Cool reaction toroom temperature, dilute with ethyl acetate, and filter through a shortplug of celite with additional ethyl acetate. Wash organics with water,dry over MgSO₄, filter, and evaporate. Purification by flash columnchromatography yields 2′,4′-difluorobiphenyl-4-carboxylic acid methylester as a yellow solid. Dissolve the purified ester in dioxane (5 mL)and add 5M NaOH (1 mL). Stir vigorously at 50° C. for 15 hours. Uponcompletion, acidify the reaction with conc. HCl and extract with ethylacetate. Evaporation of the solvent yields 300 mg (24.7%) of the titlecompound. MS (ES): m/z 233.0 (M−H).

Method F 6-(2,6-Difluorophenyl)pyridine-3-carboxylic acid

Dissolve 6-chloropyridine-3-carboxylic acid methyl ester (6.86 g, 40mmol) in toluene (100 mL) and heat to 90° C. Add phosphorous oxybromide(25 g, 87 mmol) in several portions and continue heating for 3 hours.Cool the reaction to room temperature and pour onto ice water. Extractthe reaction with ethyl acetate and wash organics again with water thenNaHCO₃. Combine organics, dry over MgSO₄, filter, and evaporate toorange solid (8.1 g, 94%) which is an 8:1 mixture of6-bromopyridine-3-carboxylic acid methylester:6-chloromopyridine-3-carboxylic acid methyl ester by ¹H NMR.

Combine the mixture as obtained above (0.225 g, 1.04 mmol) withhexamethylditin (0.375 g, 1.15 mmol), Pd(OAc)₂ (21 mg, 0.09 mmol), andtriphenylphosphine (25 mg, 0.09 mmol) in toluene (5 mL). Purge with N₂and stir at 80° C. for 18 hours. Cool reaction to room temperature. Adda solution of 1-bromo-2,6-difluorobenzene (250 mg, 1.29 mmol) in toluene(1 mL) followed by Pd(OAc)₂ (21 mg, 0.09 mmol) and triphenylphosphine(25 mg, 0.09 mmol). Purge with N₂ and stir at 80° C. for an additional18 hours. Cool reaction to room temperature. Evaporate the solvent andpurify by column chromatography (silica, 10% ethyl acetate in hexane) togive 50 mg (20% yield) of 6-(2,6-difluorophenyl)pyridine-3-carboxylicacid ethyl ester. Hydrolyze the ester with 1N sodium hydroxide solution(0.22 mL, 0.22 mmol) in methanol (3 mL) at room temperature for 3 days.Remove the volatiles under vacuum and combine the residue with 1Nhydrochloric acid solution. Collect the white solid by filtration, washwith water, and dry under vacuum to give 30 mg (63% yield) of the titlecompound. MS (ES): m/z 235.9 (M+H).

Method G 3-Fluorobiphenyl-4-carboxylic acid

Combine methyl 2-fluoro-4-bromobenzoate (1.25 g, 5.36 mmol),phenylboronic acid (1.30 g, 10.72 mmol) and CsF (2.02 g, 13.40 mmol) inDMF (25 mL) and water (3.0 mL) with stirring. Place the hetereogeneousreaction mixture open to the air in an oil bath maintained at 80° C.After 5 minutes of heating, add Pd(OAc)₂ (120 mg, 0.536 mmol) in oneportion and stir until reaction turns black. Cool reaction to roomtemperature, dilute with ethyl acetate and filter through a short plugof celite with additional ethyl acetate. Wash organics with water, dryover MgSO₄, filter and evaporate. Purification by flash columnchromatography yields 3-fluorobiphenyl-4-carboxylic acid methyl ester asa solid. Dissolve the purified ester in THF (0.25M) and add an equalvolume of 1M NaOH. Stir vigorously at room temperature for 15 hours.Upon completion, acidify the reaction with conc. HCl and extract withethyl acetate. Evaporation of the solvent yields 965 mg (84%) of thetitle compound. MS (ES): m/z 214.9 (M−H).

The following compounds are prepared essentially as described above.

3-Fluoro-2′-methylbiphenyl-4-carboxylic MS (ES): m/z 229.0(M − H) acid2′-Chloro-3-fluorobiphenyl-4-carboxylic MS (ES): m/z 205.1(M − H) acid3-Fluoro-2′-trifluoromethylbiphenyl-4- MS (ES): m/z 283.1(M − H)carboxylic acid

Method H 2-Fluoro-6-phenylpyridine-3-carboxylic acid

Dissolve 2,6-difluoropyridine (5.0 mL, 5.51 mmol) in anhydrous THF (30mL) and cool to −40° C. Add a solution of phenyl lithium (1.8 M hexanes,30.6 mL) dropwise over 5 minutes. Stir the resulting purple reaction at−40° C. for 30 minutes and bring to room temperature. Quench thereaction with water and extract the solution with ethyl acetate severaltimes. Combine the organic extracts, dry over MgSO₄, filter andevaporate onto silica gel. Purification by flash column chromatographyyields 2-fluoro-6-phenylpyridine 1.0 g (12%) as a yellow oil.

Cool a solution of LDA (3.46 mmol) in anhydrous THF (6 mL) to −78° C.Cannulate the 2-fluoro-6-phenylpyridine in anhydrous THF (6 mL) to thecooled LDA solution. Stir at −78° C. for 30 minutes then bubble carbondioxide gas through the solution for 10 minutes. Allow the reaction tocome to room temperature and purge with argon. Extract the reaction with1 M NaOH and discard the organics. Acidify the aqueous layer with conc.HCl and extract with ethyl acetate. Dry the organic layer over MgSO₄,filter and evaporate to yield the title compound as a light yellow solid(405 mg, 65%). MS (ES): m/z 216.1 (M−H).

Method I 3,5-Difluorobiphenyl-4-carboxylic acid

Combine 1-bromo-3,5-difluorobenzene (0.863 mL, 7.50 mmol) andphenylboronic acid (1.22 g, 10.00 mmol) and subject to conditionsdescribed in Method G to yield 1.3 g of 3,5-difluorobiphenyl.

Dissolve crude 3,5-difluorobiphenyl (1.3 g, 6.83 mmol) in THF (14 mL)and cool to −78° C. Prepare LiTMP from the addition of BuLi (1.6 M solnin hexanes, 5.33 mL) to tetramethyl piperidine (1.4 mL, 1.25 equiv) at−78° C. in THF (14 mL). Cannulate the cooled LiTMP into the cooled3,5-difluorobiphenyl and stir the reaction at −78° C. for 1 hour. Bubblecarbon dioxide gas through the solution for 5 minutes, warn the reactionto room temperature, pour into 50 mL of 1M NaOH, and extract with 50 mLEtOAc. Discard the organic layer. Acidify the remaining aqueous layerwith conc. HCl and extract twice with EtOAc. Dry the organics overMgSO₄, filter, and evaporate to give 1.22 g of the title compound as awhite solid (77%). MS (ES): m/z 233.1 (M−H).

Method J 3,2′,6′-Trifluorobiphenyl-4-carboxylic acid

Combine methyl 4-bromo-2-fluorobenzoate (3.66 g, 15.75 mmol),4,4,5,5,4′,4′,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolanyl (5.0 g,19.68 mmol) and potassium acetate (4.63 g, 47.19 mmol) in DMSO (40 mL)and purge the solution with argon. AddPdCl₂(1,1′-bis(diphenylphosphino)ferrocene)₂ (10 mol %, 1.35 g) andpurge the solution with argon again. Heat the reaction to 80° C. for 3hours and cool to room temperature. Wash the reaction with water andextract with ethyl acetate and concentrate. Redissolve the resultingblack oil in 1:2 ethyl acetate:hexanes, filter through a short plug ofsilica gel, and concentrate to get2-Fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acidmethyl ester as a yellow oil.

Dissolve the resulting yellow oil in acetone (100 mL) and combine withNaIO₄ (10.1 g, 47.25 mmol), NH₄OAc (3.63 g, 47.25 mmol), and water (50mL) at room temperature. Stir at room temperature for 18 hours, transferto a separatory funnel and extract with ethyl acetate several times. Drythe combined organics over MgSO₄, filter and concentrate to yield 3.0 gof 3-fluoro-4-carbomethoxybenzene boronic acid as an off-white solid.

The boronic acid obtained above (800 mg, 4.04 mmol) and2,6-difluorobromobenzene (1.17 g, 6.06 mmol) are coupled according tothe procedure described in Method G to give 380 mg of the titlecompound. MS (ES): m/z 251.1 (M−H).

Method K 6-Phenylpyridazine-3-carboxylic acid

Dissolve 6-Phenylpyridazin-3-ol (5.0 g, 29.06 mmol) in toluene (100 mL)and heat to 90° C. Add phosphorous oxybromide (25 g, 87.19 mmol) inseveral portions and heat the reaction for 30 minutes. Cool theresulting yellow solution to room temperature, pour onto ice water, andextract with ethyl acetate. Further wash the organic layers with waterand 1 M NaOH, dry over MgSO₄, filter, and evaporate to a yellow solid.Recrystallization from CHCl₃ gives 2.17 g of 3-bromo-6-phenylpyridazine.

Combine 3-Bromo-6-phenylpyridazine (1.0 g, 4.25 mmol) with DMF (5 mL),MeOH (5 mL), triethylamine (1.18 mL, 8.50 mmol), and Pd(OAc)₂ (76 mg,0.33 mmol) and evacuate the mixture. Add1,1′-bis(diphenylphosphino)ferrocene (235 mg, 0.42 mmol) and againevacuate the reaction. Bubble carbon dioxide gas through the solutionfor 5 minutes, and place the reaction under 50 psi (345 kPa) of carbondioxide. Heat the resulting solution at 50° C. for 18 hours. Cool thereaction to room temperature, diluted with water, and extract with ethylacetate. Dry the organics over MgSO₄, filter, and evaporate onto silicagel and subject to flash column chromatography.

Hydrolysis using conditions outlined in Method A gives 80 mg of thetitle compound. ¹H NMR (CDCl₃): 8.24 (d, 1H, J=8.8 Hz), 8.18-8.15 (m,2H), 8.0 (d, 1H, J=9.2 Hz), 7.56-7.55 (m, 3H).

Method L 6-(4-Fluorophenyl)pyridine-3-carboxylic acid

Combine 6-bromopyridine-3-carboxylic acid methyl ester (1.03 g, 4.78mmol), 4-fluorophenylboronic acid (1.88 g, 13.41 mmol), and cesiumfluoride (2.55 g, 16.78 mmol) in DMF (25 mL) and water (4 mL) withstirring. Place the hetereogeneous reaction mixture, open to the air, inan oil bath maintained at 80° C. After 5 minutes of heating, addPd(OAc)₂ (150 mg, 0.67 mmol) in one portion. After 17 hours, cool thereaction to room temperature, dilute with ethyl acetate and filterthrough a short plug of celite with additional ethyl acetate. Wash theorganics with water, dry over MgSO₄, filter and evaporate. Purificationby flash column chromatography yields6-(4-fluorophenyl)pyridine-3-carboxylic acid methyl ester as a yellowsolid. Dissolve the purified ester in THF (0.25 M) and add an equalvolume of 1 M NaOH. Stir vigorously at room temperature for 15 hours.Upon completion, acidify the reaction with conc. HCl and collect thewhite precipitate by filtration. Drying under vacuum yields 385 mg (37%)of the title compound. MS (ES): m/z 218.1 (M+H).

The following compound is prepared essentially as described above.

6-(Thien-2-yl)pyridine-3-carboxylic acid MS (ES): m/z 205.9(M + H)

Method M 6-(4-Fluoro-2-methylphenyl)pyridine-3-carboxylic acid

Combine 6-bromopyridine-3-carboxylic acid methyl ester (387 mg, 1.79mmol), 4-fluoro-2-methylphenylboronic acid (338 mg, 2.19 mmol), Pd(OAc)₂(40 mg, 0.18 mmol), cesium fluoride (27 mg, 0.18 mmol) and sodiumcarbonate (570 mg, 5.38 mmol) in DMF (6 mL) and water (6 mL) withstirring. Purge the reaction mixture with N₂, add triphenylphosphine (47mg, 0.18 mmol), and purge again with N₂. Place the sealed reaction in anoil bath maintained at 80° C. and allow to stir for 17 hours. Cool thereaction to room temperature and pass through a short plug of silicagel. Wash the column with dichloromethane (100 mL) followed by aqueousmethanol (100 mL, 3 methanol/1 water). Reduce the combined fractions invacuo and suspend the residual solid in water (10 mL). Filter to removea black solid and acidify with 1N hydrochloric acid solution to pH 4. Awhite precipitate forms which is collected by filtration and dried togive 306 mg (74%) of the title compound. MS (ES): m/z 231.9 (M+H).

The following compounds are prepared essentially as described above.

6-(2,4-Difluorophenyl)pyridine-3- MS (ES): m/z 236.0(M + H) carboxylicacid 6-(2-Fluorophenyl)pyridine-3-carboxylic MS (ES): m/z 218.0(M + H)acid 2′-Fluorobiphenyl-4-carboxylic acid MS (ES): m/z 215.1(M − H)2′-Methylbiphenyl-4-carboxylic acid MS (ES): m/z 211.2(M − H)

Preparation 1-1 Biphenyl-4-carboxylic acid(R)-(6-(1-((4-fluorobenzyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Mix 4-fluorophenol (1.0 g, 8.9 mmol),t-butyl-N-(2(R)-hydroxyethyl)carbamate (1.4 mL, 8.9 mmol) andtriphenylphosphine (2.33 g, 8.9 mmol) in tetrahydrofuran (25 mL) andcool in an ice bath. Add a solution of diethylazodicarboxylate (1.4 mL,8.9 mmol) in tetrahydrofuran (5 mL) dropwise and stir at roomtemperature for 18 hours. Remove the solvent under vacuum. Dissolve theresidue in ether and wash with 3 portions of 1N sodium hydroxidesolution. Reduce in vacuo and slurry the oil/solid in hexane and ethylacetate. Remove the solid by filtration and reduce the filtrate invacuo. Purify the resulting oil by chromatography using 50% ethylacetate in hexane to give 0.94 g of (2-(4-fluorophenoxy)ethyl)carbamicacid tert-butyl ester as an oil. MS (ES): m/z 255.1 (M+H).

Dissolve (2-(4-fluorophenoxy)ethyl)carbamic acid tert-butyl ester (435mg, 2.28 mmol) in trifluoroacetic acid (5 mL) cooled in an ice bath andstirred for 30 minutes. Reduce the solution in vacuo and dissolve inpyridine (10 mL) along with acetic anhydride (1.1 mL, 11.0 mmol). After18 hours remove the solvent under vacuum. Dissolve the residue indichloromethane and first extract with saturated sodium bicarbonatesolution followed by a 1N HCl solution extraction. Dry and reduce theorganic layer to give 272 mg of a solid that isN-(2-(4-fluorophenoxy)ethyl)acetamide which is carried withoutpurification to the next step. Mix theN-(2-(4-fluorophenoxy)ethyl)acetamide (240 mg, 1.22 mmol) withLawesson's reagent (296 mg, 0.73 mmol) in toluene and heat at 80° C. for90 minutes. Remove the solvent in vacuo to give an oil. Dissolve the oilin ether and remove the solid precipitate by filtration. Add excessiodomethane (3 mL) and leave at room temperature for 17 hours. Reduce invacuo to give an oil. Dissolve the oil in dichloromethane and wash withsaturated sodium bicarbonate solution. Dry and reduce the solution togive 92 mg (0.405 mmol) of N-(2-(4-fluorophenoxy)ethyl)thioacetimidicacid methyl ester as an oil. Dissolve this product in dichloromethaneand add trifluoromethanesulfonic acid methyl ester (90.5 μL, 0.8 mmol).After 18 hours remove the solvent to give an oil (129 mg). Dissolve thisoil in pyridine and add (R)-(6-amino-2(R)-hydroxyindan-1-yl)carbamicacid tert-butyl ester (87 mg, 0.33 mmol). After 24 hours remove thesolvent in vacuo to give an oil. Dissolve the oil in dichloromethane andwash with saturated sodium bicarbonate solution. Dry the organic layerand reduce under vacuum to give 88.9 mg of the titled compound and useas is.

Preparations 1-2 through 1-4 are prepared essentially as in Preparation1-1 and use as is.

Prep. # Compound Name 1-2 Carbamic acid tert-butyl ester(R)-(6-(1-((2-(3,4-difluorophenoxy)ethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide 1-3 Carbamic acid tert-butyl ester(R)-(6-(1-((2-(4-fluoro-3-methylphenoxy)ethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide 1-4 Carbamic acid tert-butyl ester(R)-(6-(1-((2-(2- fluorophenoxy)ethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Preparation 2-1 Carbamic acid tert-butyl ester(R)-(6-(1-(morpholin-4-yl)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve morpholine (1 g, 11.5 mmol) in pyridine (10 mL) along withacetic anhydride (10 mL, 100.0 mmol). After 4 hours remove the solventunder vacuum. Dissolve the residue in dichloromethane and first extractwith saturated sodium bicarbonate solution followed by a 1N HCl solutionextraction. Dry and reduce the organic layer to give 355 mg of an oilthat is 1-morpholin-4-yl-ethanone which is carried without purificationto the next step. Mix the 1-morpholin-4-yl-ethanone (347 mg, 2.7 mmol)with Lawesson's reagent (652 mg, 1.6 mmol) in toluene and heat at 80° C.for 4 hours. Remove the solvent in vacuo to give an oil. Dissolve theoil in ether and remove the solid precipitate by filtration. Add excessiodomethane (3 mL) and leave at room temperature for 17 hours. Collectthe precipitate by filtration yielding 365 mg of solid. Combine thissolid with (R)-(6-amino-2(R)-hydroxyindan-1-yl)carbamic acid tert-butylester (317 mg, 1.2 mmol) in pyridine (10 mL). After 24 hours remove thesolvent in vacuo to give an oil. Dissolve the oil in dichloromethane andwash with saturated sodium bicarbonate solution. Dry the organic layerand reduce under vacuum to give a residue that is purified bychromatography eluting with 2% methanol in chloroform to give 224 mg ofthe titled compound. ¹H-NMR (CDCl₃) δ 7.05 (1H, d), 6.55 (2H, m), 5.04(1H, d), 4.85 (1H, t), 4.40 (1H, q), 4.28 (1H, s), 3.75 (4H, m), 3.45(4H, m), 3.22 (1H, dd), 2.95 (1H, dd), 1.84 (3H, s), 1.45 (9H, s).

Preparation 3-1 Carbamic acid tert-butyl ester(R)-(6-(1-((2-methoxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve N-(2-methoxyethyl)methylamine (5.81 g, 65.18 mmol) in 40 mL ofpyridine at 0° C. and add acetic anhydride dropwise. After 30 minutesraise the reaction to room temperature and stir for 24 hours. Remove thesolvent in vacuo and dissolve the residue in methylene chloride and washwith 5N HCl. Dry the organic layer with magnesium sulfate. Filter andremove the solvent in vacuo to give 4.23 g of crudeN-(2-methoxyethyl)-N-methylacetamide. Dissolve this crude material (4.23g, 32.25 mmol) in 50 mL of toluene and add Lawesson's reagent (6.5197 g,16.12 mmol) to the mixture. Heat the reaction to 75° C. and allow it tostir for 18 hours. Remove the solvent in vacuo. Triturate the residuewith a 1:1 mixture of diethyl ether/pentane three times decantingcarefully from the residual solids. Combine the decanted layers andremove the solvent in vacuo to afford 2.13 g of crudeN-(2-methoxyethyl)-N-methylthioacetamide. Dissolve this crude material(2.13 g, 14.47 mmol) in 50 mL diethyl ether and add methyltrifluoromethanesulfonate (2.49 g, 15.19 mmol) and allow the reaction tostir for 21 hours. Decant the diethyl ether from the oil and trituratethe oil with diethyl ether (3×). Remove any excess diethylether from theoily residue in vacuo to obtain 2.92 g of crude triflic acid salt of(2-methoxyethyl)methyl(1-methylsulfanylethyl)amine as a black oil.Dissolve this black oil (2.92 g, 9.38 mmol) in 50 mL of pyridine and add(R)-(6-amino-2(R)-hydroxyindan-1-yl)carbamic acid tert-butyl ester (2.46g, 9.31 mmol) and allow the reaction to stir at room temperature for 23hours. Remove the solvent in vacuo and dissolve the residue in methylenechloride and wash with saturated aqueous sodium hydrogencarbonate. Drythe organic layer with magnesium sulfate. Filter and remove the solventin vacuo. Purify the residue by Biotage chromatography (10% MeOH/EtOActo 25% MeOH/EtOAc) to afford 1.304 g of the titled product (37% yield).MS (ES): m/z 378.2 (M+H).

Preparation 3-2 is prepared essentially as Preparation 3-1.

Prep. # Compound Name MS (ES): m/z 3-2 Carbamic acid tert-butyl ester(R)-(6-(1-((2- 406.3(M + H) propoxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

EXAMPLE 1-1 Biphenyl-4-carboxylic acid(R)-(6-(1-((2-methoxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve carbamic acid tert-butyl ester(R)-(6-(1-((2-methoxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide(153.6 mg, 0.407 mmol) in excess TFA at 0° C. and allow to stir for 1hour. Remove the solvent in vacuo. Dissolve the residue in 10 mLmethylene chloride and add triethylamine (411.8 mg, 4.07 mmol) andbiphenyl-4-carboxylic acid 2,5-dioxopyrrolidin-1-yl ester (118.3 mg,0.400 mmol). Allow the reaction mixture to stir for 21 hours. Dilute thereaction mixture with methylene chloride and wash it with saturatedaqueous sodium hydrogen carbonate (1×) and brine (1×). Dry the organiclayer with magnesium sulfate. Filter and remove the solvent in vacuo togive 162.1 mg of crude product. Purify via Biotage chromatography (30%MeOH/EtOAc) to afford 93.1 mg of the titled product as an oil (50%yield). MS (ES): m/z 458.3(M+H).

Examples 1-2 through 1-14 are prepared essentially as Example 1-1.

MS (ES): Ex. # Compound Name m/z 1-2 Biphenyl-4-carboxylic acid(R)-(6-(1-((2- 486.3 propoxyethyl)methylamino)ethylideneamino)-2(R)-(M + H) hydroxyindan-1-yl)amide 1-3 2′-Fluorobiphenyl-4-carboxylic acid(R)-(6-(1-((2- 475.1 methoxyethyl)methylamino)ethylideneamino)-2(R)-(M + H) hydroxyindan-1-yl)amide 1-4 3-Fluorobiphenyl-4-carboxylic acid(R)-(6-(1- 476.4 ((2-methoxyethyl)methylamino)ethylideneamino)- (M + H)2(R)-hydroxyindan-1-yl)amide 1-5 3,2′-Difluorobiphenyl-4-carboxylic acid(R)-(6-(1-((2- 494.2 methoxyethyl)methylamino)ethylideneamino)-2(R)-(M + H) hydroxyindan-1-yl)amide 1-63,2′,6′-Trifluorobiphenyl-4-carboxylic acid (R)-(6-(1- 512.2((2-methoxyethyl)methylamino)ethylideneamino)- (M + H)2(R)-hydroxyindan-1-yl)amide 1-7 3,2′-Difluorobiphenyl-4-carboxylic acid(R)-(6-(1- 492.3 (morpholin-4-yl)ethylideneamino)-2(R)-hydroxyindan-(M + H) 1-yl]amide 1-8 Biphenyl-4-carboxylic acid (R)-(6-(1-((2-(4-538.4 fluorophenoxy)ethyl)methylamino)ethylideneamino)- (M + H)2(R)-hydroxyindan-1-yl)amide 1-9 4′-Fluorobiphenyl-4-carboxylic acid(R)-(6-(1-((2-(4- 556.4fluorophenoxy)ethyl)methylamino)ethylideneamino)- (M + H)2(R)-hydroxyindan-1-yl)amide 1-10 2′,6′-Difluorobiphenyl-4-carboxylicacid (R)-(6-(1- 574.4 ((2-(4- (M + H)fluorophenoxy)ethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide 1-11 Biphenyl-4-carboxylic acid(R)-(6-(1-((2- 556.4 (3,4- (M + H)difluorophenoxy)ethyl]methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide 1-12 3-Fluorobiphenyl-4-carboxylic acid(R)-(6-(1-((2-(3,4- 574.2difluorophenoxy)ethyl)methylamino)ethylideneamino)- (M + H)2(R)-hydroxyindan-1-yl)amide 1-13 Biphenyl-4-carboxylic acid(R)-(6-(1-((2-(4-fluoro-3- 552.5methylphenoxy)ethyl)methylamino)ethylideneamino)- (M + H)2(R)-hydroxyindan-1-yl)amide 1-14 Biphenyl-4-carboxylic acid(R)-(6-(1-((2-(2- 538.3fluorophenoxy)ethyl)methylamino)ethylideneamino)- (M + H)2(R)-hydroxyindan-1-yl)amide

EXAMPLE 2-1 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((2-methoxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

To a mixture of thioacetamide (10.04 g, 133.6 mmol) and potassiumcarbonate (45.80 g, 331.4 mmol) in 100 mL tetrahydrofuran at 0° C. addphthaloyl chloride (28.49 g, 140.3 mmol) dropwise. Raise the reactiontemperature to 25° C. after 2 hours and allow it to stir for anadditional 2 hours before cooling the reaction mixture to 0° C. again.Quench the reaction by adding 125 mL of ice water dropwise. Extract thereaction mixture with EtOAc (2×). Dry the organic layer with magnesiumsulfate and remove the solvent in vacuo to yield 3.7 g of2-thioacetylisoindole-1,3-dione as a crude reddish solid. See also J.Org. Chem. 1997, 62, 3808-3809. ¹H NMR (CDCl3) δ 8.00 (2 H, m), 7.82 (2H, m), 3.10 (3 H, s).

Dissolve N-(2-methoxymethyl)methylamine (101.0 mg, 1.13 mmol) in 20 mLdiethyl ether at 25° C. To this add 2-thioacetyl-isoindole-1,3-dione(243.0 mg, 1.18 mmol) and allow to stir for 22 hours. Filter thereaction mixture and add methyl trifluoromethanesulfonate (194.7 mg,1.19 mmol) to the filtrate and allow to stir for an additional 18 hours.Decant the diethyl ether from the oil and triturate the oil with diethylether (3×). Remove any excess diethyl ether from the oily residue invacuo to obtain 320.4 mg of crude triflic acid salt of(2-methoxyethyl)methyl(1-methylsulfanylethyl)amine as an oil. Dissolvethis crude product (161.1 mg, 0.483 mmol) in 5 mL pyridine and addN-(R)-(6-amino-2(R)hydroxyindan-1-yl)-4-bromobenzamide (115.1 mg, 0.331mmol). Allow the reaction to stir at 25° C. for 22 hours. Remove thesolvent in vacuo and partition the residue between methylene chlorideand saturated aqueous sodium hydrogencarbonate. Dry the organic layerwith magnesium sulfate. Filter and remove the solvent in vacuo to give40.8 mg of crude product. Purify via silica gel chromatography (5%MeOH/CHCl₃ to 30% MeOH/CHCl₃) to afford 40.8 mg of the titled product(27% yield). MS (ES): m/z 460.2(M+H).

Examples 2-2 and 2-3 are prepared essentially as Example 2-1.

Ex. # Compound Name MS (ES): m/z 2-2 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((2- 446.1 (M + H) methoxyethyl)amino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide 2-3 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((2- 460.2 (M + H) methoxypropyl)amino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

EXAMPLE 3-1 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((2-methoxypropyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve 3-methoxypropylamine (154.1 mg, 1.73 mmol) in 20 mL diethylether at 25° C. To this add 2-thioacetylisoindole-1,3-dione (370.7 mg,1.81 mmol) and allow to stir for 18 hours. Filter the reaction mixtureand add methyl trifluoromethanesulfonate (298.1 mg, 1.82 mmol) to thefiltrate and allow to stir for an additional 24 hours. Decant thediethyl ether from the oil and triturate the oil with diethyl ether(3×). Remove any excess diethyl ether from the oily residue in vacuo toobtain 320.4 mg of crude triflic acid salt ofN-(3-methoxypropyl)thioacetimidic acid methyl ester as an oil. Dissolvethis crude product (379.8 mg, 1.22 mmol) in methylene chloride and washit with saturated aqueous sodium hydrogen carbonate. Dry the organiclayer with magnesium sulfate. Add methyl trifluoromethanesulfonate(210.2 mg, 1.28 mmol) to the filtrate and allow to stir for anadditional 24 hours. Remove the solvent in vacuo and triturate the oilwith diethyl ether (3×). Remove any excess diethyl ether from the oilyresidue in vacuo to obtain 296.5 mg of crude triflic acid salt of(3-methoxypropyl)methyl(1-methylsulfanylethyl)amine as an oil. Dissolvethis crude material (99.5 mg, 0.30 mmol) in 5 mL pyridine and addN-(R)-(6-amino-2(R)-hydroxyindan-1-yl)-4-bromobenzamide (105.7 mg, 0.304mmol). Allow the reaction to stir at 25° C. for 18 hours. Remove thesolvent in vacuo and partition the residue between methylene chlorideand saturated aqueous sodium hydrogen carbonate. Dry the organic layerwith magnesium sulfate. Filter and remove the solvent in vacuo to give40.8 mg of crude product. Purify via silica gel chromatography (5%MeOH/CHCl₃ to 30% MeOH/CHCl₃) to afford 50.0 mg of the titled product(35% yield). MS (ES): m/z 474.1 (M+H).

Examples 3-2 through 3-7 are prepared essentially as Example 3-1.

Ex. # Compound Name MS (ES): m/z 3-2 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((2- 522.2 (M + H) phenoxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide 3-3 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((2- 474.1 (M + H)ethoxyethyl)methylamino)ethylideneamino)-2(R)- hydroxyindan-1-yl)amide3-4 4-Bromophenyl-1-carboxylic acid (R)-(6-(1-((2- 488.1 (M + H)propoxyethyl)methylamino)ethylideneamino)- 2(R)-hydroxyindan-1-yl)amide3-5 4-Bromophenyl-1-carboxylic acid (R)- 474.1 (M + H)(6-(1-((2-methoxy-1- methylethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide 3-6 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((2- 536.2 (M + H)benzyloxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide 3-7 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((2- 488.2 (M + H)isopropoxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

EXAMPLE 4-1 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-(((R)-2-methoxy-2-phenylethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve (R)-(−)-2-methoxy-2-phenylethanol (1.1177 g, 7.34 mmol) andtriethylamine (1.11 g, 11.02 mmol) in 20 mL methylene chloride at 0° C.Dissolve para-toluenesolfonyl chloride (1.4498 g, 7.60 mmol) in 10 mLmethylene chloride and add it dropwise to the reaction. After 30 minutesraise the reaction to room temperature and allow it to stir for 24hours. Dilute the reaction with methylene chloride and wash it withwater (1×), 1N HCl (1×), saturated aqueous sodium hydrogen carbonate(1×) and brine (1×). Dry the organic layer with magnesium sulfate.Filter and remove the solvent in vacuo to give 2.01 g of crudetoluene-4-sulfonic acid 2-methoxy-2-phenylethyl ester. Dissolve thiscrude material (2.01 g, 6.56 mmol) and di-tert-butyl iminodicarboxylate(1.495 g, 6.88 mmol) in 25 mL acetonitrile. Add potassium tert-butoxide(777.1 mg, 6.92 mmol) to the reaction mixture and heat it to reflux for24 hours. Cool the reaction to room temperature and remove the solventin vacuo. Dissolve the residue in methylene chloride and wash it withwater (1×) and brine (1×). Dry the organic layer with magnesium sulfate.Filter and remove the solvent in vacuo to give 2.34 g of crude material.Purify via Biotage chromatography (5% EtOAc/hexanes) to afford 607.9 mgof (2-methoxy-2-phenylethyl)dicarbamic acid tert-butyl ester as an oil(26% yield). MS (ES): m/z 458.3(M+H).

Dissolve (2-methoxy-2-phenylethyl)dicarbamic acid tert-butyl ester(607.9 mg, 1.73 mmol) in excess trifluoroacetic acid at 0° C. and allowto stir for 1 hour. Remove the solvent in vacuo to afford crude2-methoxy-2-phenylethylamine. Dissolve this crude material in 10 mL ofpyridine at 0° C. and add acetic anhydride (194.2 mg, 1.90 mmol)dropwise. After 30 minutes raise the reaction to room temperature andstir for 24 hours. Remove the solvent in vacuo and dissolve the residuein methylene chloride and wash with 5N HCl. Dry the organic layer withmagnesium sulfate. Filter and remove the solvent in vacuo to give 384.1mg of crude N-(2-methoxy-2-phenylethyl)acetamide. Dissolve this crudematerial (384.1 mg, 1.99 mmol) in 20 mL of toluene and add Lawesson'sreagent (404.47 mg, 1.01 mmol) to the mixture. Heat the reaction to 75°C. and allow it to stir for 22 hours. Remove the solvent in vacuo.Triturate the residue with a 1:1 mixture of diethyl ether/pentane threetimes decanting carefully from the residual solids. Combine the decantedlayers and remove the solvent in vacuo to afford 125.5 mg of crudeN-(2-methoxy-2-phenylethyl)thioacetamide. Dissolve this crude material(125.5 mg, 0.600 mmol) in 10 mL methylene chloride and add methyltrifluoromethanesulfonate (108.2 mg, 0.660 mmol) and allow the reactionto stir for 21 hours. Decant the diethyl ether from the oil andtriturate the oil with diethyl ether (2×). Remove any excess diethylether from the oily residue in vacuo to obtain 102.3 mg of crude triflicacid salt of N-(2-methoxy-2-phenylethyl)thioacetimidic acid methylester. Dissolve this crude product (102.3 mg, 0.274 mmol) in methylenechloride and wash it with saturated aqueous sodium hydrogen carbonate.Dry the organic layer with magnesium sulfate. Add methyltrifluoromethanesulfonate (56.19 mg, 0.342 mmol) to the filtrate andallow to stir for an additional 24 hours. Remove the solvent in vacuoand triturate the oil with diethyl ether (3×). Remove any excess diethylether from the oily residue in vacuo to obtain 75.0 mg of crude triflicacid salt of (2-methoxy-2-phenylethyl)methyl(1-methylsulfanylethyl)amineas an oil. Dissolve this crude material (75.0 mg, 0.194 mmol) in 10 mLpyridine and add N-(R)-(6-amino-2(R)-hydroxyindan-1-yl)-4-bromobenzamide(66.0 mg, 0.190 mmol). Allow the reaction to stir at 25° C. for 18hours. Remove the solvent in vacuo and partition the residue betweenmethylene chloride and saturated aqueous sodium hydrogen carbonate. Drythe organic layer with magnesium sulfate. Filter and remove the solventin vacuo to give 70.0 mg of crude product. Purify via Biotagechromatography (2% MeOH/EtOAc) to afford 38.6 mg of the titled product(38% yield). MS (ES): m/z 538.2 (M+H).

Examples 4-2 through 4-6 are prepared essentially as Examples 4-1 usingthe appropriate commercially available alcohol.

Ex. # Compound Name MS (ES): m/z 4-2 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((3- 488.3 (M + H) methoxybutyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl) amide 4-3 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((2- 531.3 (M + H) hexyloxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide 4-4 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((4- 564.1 (M + H)benzyloxybutyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide 4-5 4-Bromophenyl-1-carboxylic acid (R)-502.1 (M + H) (6-(1-((3-methoxy-3-methylbutyl)methylamino)ethylideneamino)-2(R)- hydroxyindan-1-yl)amide4-6 4-Bromophenyl-1-carboxylic acid (R)-(6-(1-((2- 503.1 (M + H)isobutoxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

EXAMPLE 5-1 Biphenyl-1-carboxylic acid(R)-(6-(1-((2-pentoxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve the crude triflic acid salt of(1-methylsulfanylethyl)(2-pentoxyethyl)methylamine (192.1 mg, 0.523mmol) and biphenyl-4-carboxylic acid(R)-(6-amino-2(R)-hydroxyindan-1-yl)amide (170.8 mg, 0.496 mmol) in 10mL pyridine and allow it to stir for 22 hours. Remove the solvent invacuo and partition the residue between methylene chloride and saturatedaqueous sodium hydrogen carbonate. Dry the organic layer with magnesiumsulfate. Filter and remove the solvent in vacuo to give 112.3 mg ofcrude product. Purify via Biotage chromatography (10% MeOH/EtOAc 25%MeOH/EtOAc) to afford 27.8 mg of the titled product (11% yield). MS(ES): m/z 514.5 (M+H).

Examples 5-2 and 5-3 are prepared essentially as Example 5-1.

Ex. # Compound Name MS (ES): m/z 5-2 Biphenyl-1-carboxylic acid(R)-(6-(1-((2- 500.5 butoxyethyl)methylamino)ethylideneamino)-2(R)- (M +H) hydroxyindan-1-yl)amide 5-3 Biphenyl-4-carboxylic acid (R)-(6-(1-((2-526.3 cyclohexyloxyethyl)methylamino)ethylideneamino)- (M + H)2(R)-hydroxyindan-1-yl)amide

EXAMPLE 6-1 Biphenyl-4-carboxylic acid(R)-(6-(1-((2-methylsulfanylethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve 2-(methylthio)ethylamine (5.6601 g, 62.08 mmol) in 20 mLpyridine at 0° C. and add acetic anhydride (31.69 g, 310.38 mmol)dropwise. After 30 minutes raise the reaction to room temperature andstir for 21 hours. Remove the solvent in vacuo and dissolve the residuein methylene chloride and wash with 5N HCl. Dry the organic layer withmagnesium sulfate. Filter and remove the solvent in vacuo to give 4.174g of crude N-(2-methylsulfanylethyl)acetamide. Dissolve the crudematerial (2.032 g, 15.25 mmol) in 10 mL DMF and add it dropwise to asuspension of sodium hydride (404.34 mg, 16.85 mmol) in 10 mL DMF. Allowthe reaction to stir for 24 hours at room temperature. Quench thereaction with MeI (2.598 g, 18.30 mmol). Partition the reaction mixturebetween EtOAc and brine and wash the EtOAc layer with brine (2×). Drythe organic layer with magnesium sulfate. Filter and remove the solventin vacuo to give 948.9 mg of very crudeN-methyl-N-(2-methylsulfanylethyl)acetamide. Dissolve this crudematerial (299.4 mg, 2.03 mmol) in 10 mL of toluene and add Lawesson'sreagent (417.2 g, 1.03 mmol) to the mixture. Heat the reaction to 75° C.and allow it to stir for 27 hours. Remove the solvent in vacuo.Triturate the residue with diethyl ether three times decanting carefullyfrom the residual solids. Combine the decanted layers and remove thesolvent in vacuo to afford 491.7 mg of crudeN-methyl-N-(2-methylsulfanylethyl)thioacetamide. Dissolve this crudematerial (491.7 mg, 3.01 mmol) in 10 mL diethyl ether and add MeI(316.95 mg, 2.23 mmol) and allow the reaction to stir for 21 hours.Decant the diethyl ether from the oil and triturate the oil with diethylether (3×). Remove any excess diethyl ether from the oily residue invacuo to obtain 165.6 mg of crude HI salt ofmethyl-(2-methylsulfanylethyl)(1-methylsulfanylethyl)amine as a yellowsolid. Dissolve this crude material (165.6 mg, 0.542 mmol) in 10 mL ofpyridine and add (R)-(6-amino-2(R)-hydroxyindan-1-yl)carbamic acidtert-butyl ester (197.6 mg, 0.748 mmol) and allow the reaction to stirat room temperature for 22 hours. Remove the solvent in vacuo anddissolve the residue in methylene chloride and wash with saturatedaqueous sodium hydrogen carbonate. Dry the organic layer with magnesiumsulfate. Filter and remove the solvent in vacuo. Purify the residue byBiotage chromatography (5% MeOH/EtOAc to 20% MeOH/EtOAc) to afford 81.6mg of(R)-6-(1-(methyl-(2-methylsulfanylethyl)amino)ethylideneamino)-2(R)-hydroxyindan-1-yl)carbamicacid tert-butyl ester (28%). MS (ES): m/z394.1(M+H). Prepare the finalproduct as in Example 1-1 to afford 82.4 mg of crude material. Purifyvia silica gel column chromatography (2% MeOH/CHCl₃ to 5% MeOH/CHCl₃) toafford 50.9 mg of the titled product as an oil (52% yield). MS (ES): m/z474.2(M+H).

EXAMPLE 7-1 3,2′-Difluorobiphenyl-4-carboxylic acid(R)-(6-(1-((2-methylsulfanylethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve the crude HI salt ofmethyl-(2-methylsulfanylethyl)(1-methylsulfanylethyl)amine (90.1 mg,0.295 mmol) and 3,2′-difluorobiphenyl-4-carboxylic acid(R)-(6-amino-2(R)-hydroxyindan-1-yl)amide (101.3 mg, 0.266 mmol) in 10mL pyridine. Allow the reaction to stir at 25° C. for 21 hours. Removethe solvent in vacuo and partition the residue between methylenechloride and saturated aqueous sodium hydrogen carbonate. Dry theorganic layer with magnesium sulfate. Filter and remove the solvent invacuo to give 91.5 mg of crude product. Purify via silica gel columnchromatography (2% MeOH/CHCl₃ to 5% MeOH/CHCl₃) to afford 30.5 mg of thetitled product (22% yield). MS (ES): m/z 510.1(M+H).

Example 7-2 is prepared essentially as Example 7-1.

Ex. # Compound Name MS (ES): m/z 7-2 3,2′-Difluorobiphenyl-4-carboxylicacid (R)- 552.1 (6-(1-((2-tert- (M + H)butylsulfanylethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

EXAMPLE 8-1 3,2′-Difluorobiphenyl-4-carboxylic acid(R)-(6-((1-thiomorpholin-4-yl)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve crude 1-thiomorpholin-4-yl-ethanethione (869.4 mg, 5.39 mmol)in 20 mL diethyl ether and add MeI (501.0 mg, 3.53 mmol). Allow thereaction to stir at room temperature for 18 hours. Decant the diethylether from the oil and triturate the oil with diethyl ether (2×). Removeany excess diethyl ether from the oily residue in vacuo to obtain 142.2mg of crude HI salt of 4-(1-methylsulfanylethyl)thiomorpholine as asolid. Dissolve this crude material (142.2 mg, 0.469 mmol) in 20 mL ofpyridine and add (R)-(6-amino-2(R)-hydroxyindan-1-yl)carbamic acidtert-butyl ester (110.0 mg, 0.469 mmol) and allow the reaction to stirat room temperature for 22 hours. Remove the solvent in vacuo anddissolve the residue in methylene chloride and wash with saturatedaqueous sodium hydrogen carbonate. Dry the organic layer with magnesiumsulfate. Filter and remove the solvent in vacuo. Purify via silica gelcolumn chromatography (2% MeOH/CHCl₃) to afford 74.1 mg of crude(R)-6-((1-thiomorpholin-4-yl)ethylideneamino)-2(R)-hydroxyindan-1-yl)carbamicacid tert-butyl ester. Crude MS (ES): m/z392.2 (M+H). Dissolve thiscrude material (74.1 mg, 0.189 mmol) in excess TFA at 0° C. and allow tostir for 1 hour. Remove the solvent in vacuo. Dissolve the residue in 10mL methylene chloride and add triethylamine (191.5 mg, 1.89 mmol) and3,2′-difluorobiphenyl-4-carboxylic acid 2,5-dioxopyrrolidin-1-yl ester(64.3 mg, 0.194 mmol). Allow the reaction mixture to stir for 17 hours.Dilute the reaction mixture with methylene chloride and wash it withsaturated aqueous sodium hydrogen carbonate (1×) and brine (1×). Dry theorganic layer with magnesium sulfate. Filter and remove the solvent invacuo to give 73.6 mg of crude product. Purify via silica gel columnchromatography (2% MeOH/CHCl₃) to afford 3.2 mg of the titled product.(3% yield). MS (ES): m/z 508.2 (M+H).

EXAMPLE 9-1 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((2-tert-butoxyethyl)amino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve ethylene glycol mono-tert-butyl ether (2.62 g, 22.17 mmol) andtriethylamine (3.36 g, 11.02 mmol) in 40 mL methylene chloride at 0° C.Dissolve para-toluenesulfonyl chloride (4.2277 g, 22.17 mmol) in 10 mLmethylene chloride and add it dropwise to the reaction. After 30 minutesraise the reaction to room temperature and allow it to stir for 17hours. Dilute the reaction with methylene chloride and wash it withwater (1×), 1N HCl (1×), saturated aqueous sodium hydrogen carbonate(1×) and brine (1×). Dry the organic layer with magnesium sulfate.Filter and remove the solvent in vacuo to give 4.6387 g of crudetoluene-4-sulfonic acid 2-tert-butoxyethyl ester. Dissolve this crudematerial (1.2423 g, 4.56 mmol) and di-benzyl iminodicarboxylate (1.4385g, 5.04 mmol) (see Synthesis, 1988, 992-994) in 20 mL acetonitrile. Addpotassium tert-butoxide (575.9 mg, 5.13 mmol) to the reaction mixtureand heat it to reflux for 24 hours. Cool the reaction to roomtemperature and remove the solvent in vacuo. Dissolve the residue inmethylene chloride and wash it with water (1×) and brine (1×). Dry theorganic layer with magnesium sulfate. Filter and remove the solvent invacuo to give 1.9215 g of crude material. Purify via Biotagechromatography (EtOAc/Hexanes) to afford 387.8 mg of(2-tert-butoxyethyl)dicarbamic acid benzyl ester (22%). MS (ES): m/z386.3 (M+H).

Dissolve (2-tert-butoxyethyl)dicarbamic acid benzyl ester (387.8 mg,1.01 mmol) and 5% Pd/C (0.212 g) in absolute ethanol and expose it to 60psi of H₂ for 18 hours. Filter the reaction mixture over a pad ofcelite. Acidify the filtrate with 5N HCl and concentrate in vacuo toobtain 139.4 mg of a crude residue containing the HCl salt of2-tert-butoxyethylamine. Dissolve this crude material (139.4 mg, 0.852mmol) in 10 mL pyridine at 25° C. To this add2-thioacetylisoindole-1,3-dione (180.53 mg, 0.88 mmol) and allow to stirfor 23 hours. Concentrate the reaction mixture in vacuo and dissolve theresidue in methylene chloride and wash it with 1N HCl (1×). Dry theorganic layer with magnesium sulfate. Filter and remove the solvent invacuo to give 266.9 mg of crude product. Triturate this crude materialin diethyl ether and remove the solids by filtration. Concentrate thefiltrate in vacuo to afford 176.2 mg of a crude solid. Purify viaBiotage chromatography to afford 74.1 mg ofN-(2-tert-butoxyethyl)thioacetamide. Dissolve this thioacetamide (74.1mg, 0.423 mmol) in 20 mL methylene chloride and add methyltrifluoromethanesulfonate (76.3 mg, 0.465 mmol). Allow the reactionmixture to stir for an additional 21 hours. Decant the diethyl etherfrom the oil and triturate the oil with diethyl ether (3×). Remove anyexcess diethyl ether from the oily residue in vacuo to obtain 60.3 mg ofcrude triflic acid salt of N-(2-tert-butoxyethyl)thioacetimidic acidmethyl ester as an oil. Dissolve this crude product (60.3 mg, 0.62.5mmol) in 10 mL pyridine and addN-(R)-(6-amino-2(R)-hydroxyindan-1-yl)-4-bromobenzamide (62.5 mg, 0.180mmol). Allow the reaction to stir at 25° C. for 22 hours. Remove thesolvent in vacuo and partition the residue between methylene chlorideand saturated aqueous sodium hydrogen carbonate. Dry the organic layerwith magnesium sulfate. Filter and remove the solvent in vacuo to give31.4 mg of crude product. Purify via Biotage chromatography (10%MeOH/EtOAc) to afford 6.6 mg of the titled product (8% yield). MS (ES):m/z 488.1 (M+H).

EXAMPLE 10-1 Biphenyl-4-carboxylic acid(R)-(6-(1-((2-tert-butoxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve the crude HI salt of(2-tert-butoxyethyl)methyl-(1-methylsulfanylethyl)amine (obtained usingthe crude HCl salt of 2-tert-butoxyethylamine, referenced in Example9-1, using the methodology detailed in Example 6-1) (297.1 mg, 0.897mmol) in 20 mL pyridine and add biphenyl-4-carboxylic acid(R)-(6-amino-2(R)-hydroxyindan-1-yl)amide (290.7 mg, 0.844 mmol). Allowthe reaction to stir for 18 hours. Remove the solvent in vacuo anddissolve the residue in methylene chloride and wash with saturatedaqueous sodium hydrogen carbonate. Dry the organic layer with magnesiumsulfate. Filter and remove the solvent in vacuo to afford 248.7 mg ofcrude product. Purify the residue by Biotage chromatography (10%MeOH/EtOAc to 25% MeOH/EtOAc) to afford 91.7 mg of the titled product(22% yield). MS (ES): m/z 500.5 (M+H).

EXAMPLE 11-1 3,2′-Difluorobiphenyl-1-carboxylic acid(R)-(6-(1-((2-tert-butoxyethyl)amino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve the crude HI salt of N-(2-tert-butoxyethyl)thioacetimidic acidmethyl ester (see Example 9-1) (91.2 mg, 0.287 mmol) in 10 mL pyridineand add 3,2′-difluorobiphenyl-4-carboxylic acid(R)-(6-amino-2(R)-hydroxyindan-1-yl)amide (102.4 mg, 0.269 mmol). Allowthe reaction to stir for 20 hours. Remove the solvent in vacuo anddissolve the residue in methylene chloride and wash with saturatedaqueous sodium hydrogen carbonate. Dry the organic layer with magnesiumsulfate. Filter and remove the solvent in vacuo to afford 71.6 mg ofcrude product. Purify the residue by Biotage chromatography (5%MeOH/EtOAc to 20% MeOH/EtOAc) to afford 45.7 mg of the titled product(33% yield). MS (ES): m/z 522.2 (M+H).

EXAMPLE 12-1 3,2′-Difluorobiphenyl-4-carboxylic acid(R)-(6-(1-((2-(1-methylcyclopropoxy)ethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve 2-(1-methylcyclopropoxy)ethanol (362.3 mg, 3.12 mmol) (see Tet.Lett. 1999, 40, 8647-8650) and triethylamine (473.4 mg, 4.68 mmol) in 10mL methylene chloride at 0° C. Dissolve para-toluenesulfonyl chloride(596.1 mg, 3.12 mmol) in 10 mL methylene chloride and add it dropwise tothe reaction. After 30 minutes raise the reaction to room temperatureand allow it to stir for 24 hours. Dilute the reaction with methylenechloride and wash it with water (1×), 1N HCl (1×), saturated aqueoussodium hydrogen carbonate (1×) and brine (1×). Dry the organic layerwith magnesium sulfate. Filter and remove the solvent in vacuo to give670.1 mg of crude toluene-4-sulfonic acid 2-(1-methylcyclopropoxy)ethylester. Dissolve N-methylacetamide (176.9 mg, 2.42 mmol) in 5 mLtetrahydrofuran and add it dropwise to a slurry of sodium hydride (122.9mg, 3.07 mmol) in 5 mL tetrahydrofuran. Allow the reaction to stir atroom temperature for 22 hours. Dissolve the crude toluene-4-sulfonicacid 2-(1-methylcyclopropoxy)ethyl ester (670.1 mg, 2.48 mmol) in 5 mLtetrahydrofuran and add it dropwise to the reaction mixture. After theaddition is complete, heat the reaction to reflux and allow it to stirfor 26 hours. Cool the reaction and remove the solvent in vacuo.Dissolve the residue in methylene chloride and wash with brine (1×). Drythe organic layer with magnesium sulfate. Filter and remove the solventin vacuo to afford 276.9 mg of crude material. Purify the residue byBiotage chromatography (50% EtOAc/Hexanes to 30% MeOH/EtOAc) to afford83.3 mg of N-methyl-N-(2-(1-methylcyclopropoxy)ethyl)acetamide. Dissolvethis material (83.3 mg, 0.486 mmol) in 10 mL of toluene and addLawesson's reagent (102.5 mg, 0.253 mmol) to the mixture. Heat thereaction to 75° C. and allow it to stir for 24 hours. Remove the solventin vacuo. Triturate the residue with diethyl ether three times decantingcarefully from the residual solids. Combine the decanted layers andremove the solvent in vacuo to afford 47.0 mg of crudeN-methyl-N-(2-(1-methylcyclopropoxy)ethyl)thioacetamide. Dissolve thiscrude material (47.0 mg, 0.251 mmol) in 10 mL diethyl ether, add MeI(excess) and allow the reaction to stir for 21 hours. Decant the diethylether from the oil and triturate the oil with diethyl ether (2×). Removeany excess diethyl ether from the oily residue in vacuo to obtain 46.5mg of crude HI salt ofmethyl-(2-(1-methylcyclopropoxy)ethyl)-(1-methylsulfanylethyl)amine asan oil. Dissolve this crude material (46.5 mg, 0.141 mmol) in 10 mL ofpyridine and add 3,2′-difluorobiphenyl-4-carboxylic acid(R)-(6-amino-2(R)-hydroxyindan-1-yl)amide (49.6 mg, 0.130 mmol) andallow the reaction to stir at room temperature for 20 hours. Remove thesolvent in vacuo and dissolve the residue in methylene chloride and washwith saturated aqueous sodium hydrogen carbonate. Dry the organic layerwith magnesium sulfate. Filter and remove the solvent in vacuo to give39.2 mg of crude product. Purify via silica gel column chromatography(2% MeOH/CHCl₃) to afford 10.7 mg of the titled product (15% yield). MS(ES): m/z 534.2 (M+H).

EXAMPLE 13-1 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((2-methoxyethyl)methylamino)propylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve N-(2-methoxymethyl)methylamine (996.1 mg, 11.17 mmol) andtriethylamine (1.3536 g, 13.404 mmol) in 10 mL methylene chloride andadd propionic anhydride (1.526 g, 11.73 mmol) to the reaction mixture.Allow the reaction to stir at room temperature for 20 hours. Dilute thereaction mixture with methylene chloride and wash it with water (1×),saturated aqueous sodium hydrogencarbonate (1×), 1N HCl (1×) and brine(1×). Dry the organic layer with magnesium sulfate. Filter and removethe solvent in vacuo to give 854.9 mg of crudeN-(2-methoxyethyl)-N-methylpropionamide. Dissolve this material (77.8mg, 5.36 mmol) in 20 mL of toluene and add Lawesson's reagent (1.091 g,2.70 mmol) to the mixture. Heat the reaction to 75° C. and allow it tostir for 22 hours. Remove the solvent in vacuo. Triturate the residuewith diethyl ether three times decanting carefully from the residualsolids. Combine the decanted layers and remove the solvent in vacuo toafford 446.6 mg of crude N-(2-methoxyethyl)-N-methylthiopropionamide.Dissolve this crude material in 20 mL methylene chloride, add methyltrifluoromethanesulfonate (477.2 mg, 2.91 mmol) and allow the reactionto stir for 26 hours. Remove the solvent in vacuo and triturate the oilwith diethyl ether (3×). Remove any excess diethyl ether from the oilyresidue in vacuo to obtain 873.8 mg of crude triflic acid salt of(2-methoxyethyl)methyl-(1-methylsulfanylpropyl)amine as an oil. Dissolvethis crude material (131.3 mg, 0.404 mmol) in 10 mL of pyridine and add3,2′-difluorobiphenyl-4-carboxylic acid(R)-(6-amino-2(R)-hydroxyindan-1-yl)amide (111.9 mg, 0.322 mmol) andallow the reaction to stir at room temperature for 24 hours. Remove thesolvent in vacuo and dissolve the residue in methylene chloride and washwith saturated aqueous sodium hydrogen carbonate. Dry the organic layerwith magnesium sulfate. Filter and remove the solvent in vacuo to givethe crude product. Purify via Biotage chromatography (10% MeOH/EtOAc) toafford 66.1 mg of the titled product (44% yield). MS (ES): m/z 474.2(M+H).

EXAMPLE 14-1 Biphenyl-4-carboxylic acid(R)-(6-(1-((2-methylsulfanylethyl)amino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Dissolve N-(2-methylsulfanylethyl)acetamide (352.7 mg, 2.65 mmol)(seeExample 6-1) in 50 mL of toluene and add Lawesson's reagent (537.0 g,1.33 mmol) to the mixture. Heat the reaction to 75° C. and allow it tostir for 26 hours. Remove the solvent in vacuo. Triturate the residuewith diethyl ether three times decanting carefully from the residualsolids. Combine the decanted layers and remove the solvent in vacuo toafford 400.9 mg of crude N-(2-methylsulfanylethyl)thioacetamide.Dissolve this crude material (400.9 mg, 2.69 mmol) in 10 mL methylenechloride, add MeI (458.18 mg, 3.228 mmol) and allow the reaction to stirfor 24 hours. Remove the solvent in vacuo and triturate the oil withdiethyl ether (3×). Remove any excess diethyl ether from the oilyresidue in vacuo to obtain 526.8 mg of crude HI salt ofN-(2-methylsulfanylethyl)thioacetimidic acid methyl ester as an oil.Dissolve this crude material (71.1 mg, 0.244 mmol) in 10 mL of pyridineand add biphenyl-4-carboxylic acid(R)-(6-amino-2(R)-hydroxyindan-1-yl)amide (75.3 mg, 0.2.19 mmol) andallow the reaction to stir at room temperature for 22 hours. Remove thesolvent in vacuo and dissolve the residue in methylene chloride and washwith saturated aqueous sodium hydrogen carbonate. Dry the organic layerwith magnesium sulfate. Filter and remove the solvent in vacuo to give156.0 mg of crude product. Purify via Biotage chromatography (10%MeOH/EtOAc to 25% MeOH/EtOAc) to afford 62.4 mg of the titled product(62% yield). MS (ES): m/z 460.2 (M+H).

EXAMPLE 15-1 4-Bromophenyl-1-carboxylic acid(R)-(6-(1-((2-methoxy-1(R)-phenylethyl)methylamino)ethylideneamino)2(R)-hydroxyindan-1-yl)amide

Dissolve (R)-(−)-2-phenylgylcinol (1.006 g, 7.33 mmol) and triethylamine(1.781 g, 17.60 mmol) in 10 mL methylene chloride at 0° C. Add acetylchloride (1.209 g, 15.40 mmol) to the reaction mixture. After 30 minutesallow the reaction to warm to 25° C. and Allow to stir for 12 hours.Remove the solvent in vacuo and dissolve the residue in ethyl acetate.Wash the organic layer with water (1×), 1N HCl (1×) and brine (1×).Separate the organic layer and dry it over magnesium sulfate. Filter andremove the solvent in vacuo to give 1.053 g of the crude acetic acid2-acetylamino-2-phenylethyl ester in 65% yield. Dissolve this product(1.053 g, 4.76 mmol) in 20 mL methanol and add an excess of potassiumcarbonate. Allow the reaction to stir at 25° C. for 18 hours. Remove thesolvent in vacuo and triturate the residue with methanol. Filter off allsolids and wash with copious amounts of methanol. Remove the solvent invacuo to obtain 717.4 mg of the crudeN-(2-hydroxy-1-phenylethyl)acetamide as a yellow oil in 84% yield.Dissolve this crude product (717.4 mg, 4.00 mmol) in 10 mLtetrahydrofuran and add it dropwise to a suspension of sodium hydride(376.2 g, 9.405 mmol) in 20 mL tetrahydrofuran at 25° C. and allow it tostir for 18 hours. Add excess MeI to the reaction mixture and allow itto stir for and additional 26 hours. Remove the solvent in vacuo anddissolve the residue in methylene chloride. Wash the organic layer withwater (1×) and brine (1×) and dry it over magnesium sulfate. Remove thesolvent in vacuo to afford 625.2 mg of the crudeN-(2-methoxy-1-phenylethyl)-N-methylacetamide as a yellow oil. Dissolve426.1 mg of this crude product in 20 mL toluene and add Lawesson'sreagent (424.0 mg, 1.05 mmol) to the mixture. Heat the mixture to 80° C.for 21 hours. Remove the solvent in vacuo and triturate the residue witha 1:1 mixture of diethyl ether/pentane (3×). Combine the decantedorganic layers and remove the solvent in vacuo to afford 230.3 mg ofcrude titled product. Purify via column chromatography (silica gel, 15%EtOAc/Hexanes) to afford 143.3 mg ofN-(2-methoxy-1-phenylethyl)-N-methylthioacetamide as an oil in 31%yield. MS (ES): m/z 224 (M+H).

Add methyl trifluoromethanesulfonate (110.56 mg, 0.674 mmol) to asolution of N-(2-methoxy-1-phenylethyl)-N-methylthioacetamide in 10 mLdiethyl ether and allow the reaction to stir at 25° C. for 18 hours.Decant the diethyl ether from the oil and triturate the oil with diethylether (3×). Remove any excess diethyl ether from the oily residue invacuo to obtain 235.5 mg of crude thioimidate ofN-(2-methoxy-1-phenylethyl)-N-methylthioacetamide as an oil. Dissolvethis crude product (235.5 mg, 0.608 mmol) in 20 mL pyridine and addN-(R)-(6-amino-2(R)-hydroxyindan-1-yl)-4-bromobenzamide (201.7 mg, 0.581mmol). Allow the reaction to stir at 25° C. for 23 hours. Remove thesolvent in vacuo and partition the residue between methylene chlorideand saturated aqueous sodium hydrogen carbonate. Dry the organic layerwith magnesium sulfate. Filter and remove the solvent in vacuo to give257.0 mg of crude product. Purify via Biotage chromatography (5%MeOH/EtOAc) to afford 162.9 mg of the titled product as a cream coloredsolid (52% yield). MS (ES): m/z 537(M+H).

Examples 15-2 through 15-5 are prepared essentially as Example 15-1.

Ex. # Compound Name MS (ES): m/z 15-2 4-Bromophenyl-1-carboxylic acid(R)- 552.1 (M + H) (6-(1-((2-methoxy-1-phenylethyl)methylamino)ethylideneamino)-2(R)- hydroxyindan-1-yl)amide15-3 3,2′-Difluorobiphenyl-4- 536.2 (M + H) carboxylic acid(R)-(6-(1-(2- tertbutoxyethyl)methylamino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide 15-4 3,2′-Difluorobiphenyl-4- 520.0 (M + H)carboxylic acid (R)-(6-(1-((2- cyclopropoxyethyl)methyl-amino)ethylideneamino)- 2(R)-hydroxyindan-1-yl)amide 15-53,2′-Difluorobiphenyl-4- 550.2 (M + H) carboxylic acid(R)-(6-(1-((2-(1,1- dimethylpropoxy)ethyl)methyl-amino)ethylideneamino)- 2(R)-hydroxyindan-1-yl)amide

EXAMPLE 16-1 Biphenyl-4-carboxylic acid(R)-(6-(1-((2-(pyridin-2-ylsulfanylethyl)amino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

To a mixture of 2-aminoethanethiol hydrochloride (4.07 g, 35.6 mmol) indioxane (75 mL) at 50° C., add sodium hydride (60% in mineral oil, 2.84g, 71.0 mmol) at room temperature under nitrogen. Allow the reaction tostir for 5 minutes and add 2-chloropyridine (3.5 mL, 37 mmol) to themixture. Reflux the mixture for 24 hours and cool to room temperature.Add water (100 mL), and methylene chloride (300 mL) to this mixture.Extract the aqueous layer with methylene chloride (3×50 mL). Wash thecombined organic phase with brine (200 mL), dry over magnesium sulfate,filter and concentrate to an orange oil. Flash chromatography on silicagel eluting with 50% (80:18:12 CHCl₃/MeOH/concentrated NH₄OH)/methylenechloride affords the title compound as a yellow oil (1.67 g, 30%). ¹HNMR (CDCl₃) δ 8.43 (m, 1H), 7.44 (m, 1H), 7.21 (m, 1H), 6.97 (m, 1H),3.30 (m, 2H), 3.02 (m, 2H); MS (ES) m/z:═155 [C₇H₁₀N₂S+H]⁺. Add2-thioacetyl-isoindole-1,3-dione (0.7 g, 3.2 mmol) to a solution of2-(pyridin-2-ylsulfanyl)ethylamine (0.5 g, 3.2 mmol) in CHCl₃ (20 mL) at0° C. After 15 minutes, concentrate and purify the residue by flashchromatography on silica gel eluting with 40% EtOAc/Hex containing 2%NH₄OH to afford N-(2-(pyridin-2-ylsulfanyl)ethyl)thioacetamide (0.4 g,58%) as a pale yellow oil. ¹H NMR (CDCl₃) δ 10.26 (br s, 1H), 8.41-8.43(m, 1H), 7.57 (dt, J=7.35 Hz, J=1.8 Hz, 1H), 7.31 (dd, J=8.10 Hz, J=0.96Hz, 1H), 7.09-7.13 (m, 1H), 3.87-3.92 (m, 2H), 3.40-3.43 (m, 2H), 2.53(s, 3H); MS (APCI): m/z 213 (M+H).

Add methyl trifluoromethanesulfonate (65.5 mg, 0.4 mmol) to a solutionof N-(2-(pyridin-2-ylsulfanyl)ethyl)thioacetamide (85.0 mg, 0.4 mmol) inmethylene chloride (10 mL) under nitrogen. Stir the reaction mixture atroom temperature for 5 minutes and remove the solvent under reducedpressure to give N-[2-(pyridin-2-ylsulfanyl)ethyl)thioacetimidic acidmethyl ester (0.4 mmol) as a colorless oil. MS (ES): m/z 227 (M+H).

Dissolve N-[2-(pyridin-2-ylsulfanyl)ethyl)thioacetimidic acid methylester (0.4 mmol) in dry pyridine (5.0 mL) and add biphenyl-4-carboxylicacid (R)-(6-amino-2(R)-hydroxy-indan-1-yl)amide (0.15 g, 0.4 mmol). Thereaction mixture was stirred at room temperature for 12 hours. Removethe solvent under reduced pressure and purify the residue by flashchromatography on silica gel eluting with 20% MeOH/EtOAc to afford thetitle compound (140 mg, 67%) as a colorless oil. ¹H NMR (CDCl₃) δ 8.40(s, 1H), 7.89 (d, J=8.4 Hz, 2H), 7.60-7.68 (m, 4H), 7.36-7.51 (m, 4H),7.11 (d, J=7.8 Hz, 1H), 7.00 (t, J=5.4 Hz, 1H), 6.68-6.73 (m, 4H), 5.95(br s, 1H), 5.28 (s, 1H), 4.52 (q, J=7.5 Hz, 1H), 3.68 (br s, 2H), 3.39(br s, 2H), 3.30 (dd, J=15.6 Hz, J=7.8 Hz, 1 H), 2.94 (dd, J=15.3 Hz,J=7.8 Hz, 1H), 1.78 (s, 3H); MS (APCI): m/z 523 (M+H)⁺.

EXAMPLE 17-1 Biphenyl-4-carboxylic acid(R)-(6-(1-((2-(pyridin-2-ylsulfanylethyl)amino)ethylideneamino)-2(R)-hydroxyindan-1-yl)amide

Add sodium hydride (60% in mineral oil, 6.63 g, 166 mmol) to a solutionof 2-hydroxyethylamine (10.0 mL, 166 mmol) in dioxane (150 mL) at roomtemperature under nitrogen. After stirring for 10 minutes at roomtemperature, add 2-chloropyridine (15.6 mL, 166 mmol) and heat thereaction mixture to reflux. After stirring at reflux for 14 hours, coolthe reaction mixture to room temperature and dilute with water (100 mL)and methylene chloride (200 mL). Extract the aqueous layer withmethylene chloride (2×100 mL). Wash the combined organic phase withbrine (200 mL), dry over magnesium sulfate, filter, and concentrate toan orange oil. Flash chromatography on silica gel eluting with 50% of a(80:18:2 CHCl₃/MeOH/concentrated NH₄OH) solution in methylene chlorideaffords 2-phenoxy-ethylamine as a yellow oil (17.9 g, 78%). ¹H NMR(CDCl₃) δ 8.09-8.15 (m, 1H), 7.53-7.56 (m, 1H), 6.80-6.85 (m, 1H),6.70-6.75 (m, 1H), 4.27-4.31 (m, 2H), 3.06-3.10 (m, 2H); MS (ES): m/z139 (M+H).

Add 2-(pyridin-2-yloxy)ethylamine (500 mg, 3.62 mmol) to a solution of2-thioacetyl-isoindole-1,3-dione (743 mg, 3.62 mmol) in CHCl₃ (18 mL) at0° C. After stirring for 15 minutes, concentrate the solution,redissolve in ethyl acetate (25 mL), filter, and concentrate again.Flash chromatography (SiO₂, 1:1 EtOAc/Hex) givesN-(2-(pyridin-2-yloxy)ethyl)thioacetamide (350 mg, 49%) as a beige/pinksolid. ¹H NMR (CDCl₃) δ 8.79 (br s, 1H), 8.10-8.08 (m, 1H), 7.58-7.65(m, 1H), 6.92-7.00 (m, 1H), 6.76-6.82 (m, 1H), 4.57-4.65 (m, 2H),3.98-4.05 (m, 2H), 2.56 (s, 3H).

Add N-(2-(pyridin-2-yloxy)ethyl)thioacetamide (50 mg, 0.25 mmol) andmethyl trifluoromethanesulfonate (0.028 mL, 0.25 mmol) in methylenechloride (1.5 mL), and stir at room temperature. After 10 minutes,concentrate the solution to dryness. To this residue, add a solution ofbiphenyl-4-carboxylic acid (R)-(6-amino-2(R)-hydroxyindan-1-yl)amide (95mg, 0.275 mmol) in pyridine (3 mL). Stir the resulting solution at roomtemperature for 18 hours and remove the pyridine under vacuum. Flashchromatography (SiO₂, 1:1 EtOAc/Hex to 80:18:2 CHCl₃/MeOH/concentratedNH₄OH) gives the title compound (21 mg, 17%) as a white solid. ¹H NMR(CDCl₃) δ 7.88-7.95 (m, 2H), 7.65-7.74 (m, 2H), 7.60-7.70 (m, 2H),7.40-7.53 (m, 3H), 7.24-7.26 (m, 2H), 7.02-7.99 (m, 2H), 6.626.70 (m,1H), 5.35-5.42 (m, 1H), 4.55-4.62 (m, 1H), 3.78-3.90 (m, 4H), 3.30-3.43(m, 1H), 2.92-3.08 (m, 1H), 2.02 (s, 3H); MS (ES): m/z 507 (M+H).

The compounds of the present invention can be administered alone or inthe form of a pharmaceutical composition, that is, combined withpharmaceutically acceptable carriers or excipients, the proportion andnature of which are determined by the solubility and chemical propertiesof the compound selected, the chosen route of administration, andstandard pharmaceutical practice. The compounds of the presentinvention, while effective themselves, may be Formulated andadministered in the form of their pharmaceutically acceptable salts, forpurposes of stability, convenience, solubility, and the like. Inpractice, the compounds of Formula I are usually administered in theform of pharmaceutical compositions, that is, in admixture withpharmaceutically acceptable carriers or diluents.

Thus, the present invention provides pharmaceutical compositionscomprising a compound of Formula I and a pharmaceutically acceptablediluent. The present invention also provides suitable packaging,including a label, containing the pharmaceutical compositions comprisinga compound of Formula I.

The compounds of Formula I can be administered by a variety of routes.In effecting treatment of a patient afflicted with disorders describedherein, a compound of Formula I can be administered in any form or modewhich makes the compound bioavailable in an effective amount, includingoral and parenteral routes. For example, compounds of Formula I can beadministered orally, by inhalation, subcutaneously, intramuscularly,intravenously, transdermally, intranasally, rectally, occularly,topically, sublingually, buccally, and the like. Oral administration isgenerally preferred for treatment of the disorders described herein.

One skilled in the art of preparing Formulations can readily select theproper form and mode of administration depending upon the particularcharacteristics of the compound selected, the disorder or condition tobe treated, the stage of the disorder or condition, and other relevantcircumstances. (Remington's Pharmaceutical Sciences, 18th Edition, MackPublishing Co. (1990)).

The pharmaceutical compositions are prepared in a manner well known inthe pharmaceutical art. The carrier or excipient may be a solid,semi-solid, or liquid material which can serve as a vehicle or mediumfor the active ingredient. Suitable carriers or excipients are wellknown in the art. The pharmaceutical composition may be adapted fororal, inhalation, parenteral, or topical use and may be administered tothe patient in the form of tablets, capsules, aerosols, inhalants,suppositories, solutions, suspensions, or the like.

The compounds of the present invention may be administered orally, forexample, with an inert diluent or capsules or compressed into tablets.For the purpose of oral therapeutic administration, the compounds may beincorporated with excipients and used in the form of tablets, troches,capsules, elixirs, suspensions, syrups, wafers, chewing gums and thelike. These preparations should contain at least 4% of the compound ofthe present invention, the active ingredient, but may be varieddepending upon the particular form and may conveniently be between 4% toabout 70% of the weight of the unit. The amount of the compound presentin compositions is such that a suitable dosage will be obtained.Preferred compositions and preparations according to the presentinvention may be determined by a person skilled in the art.

The tablets, pills, capsules, troches, and the like may also contain oneor more of the following adjuvants: binders such as microcrystallinecellulose, gum tragacanth or gelatin; excipients such as starch orlactose, disintegrating agents such as alginic acid, Primogel, cornstarch and the like; lubricants such as magnesium stearate or Sterotex;glidants such as colloidal silicon dioxide; and sweetening agents suchas sucrose or saccharin may be added or a flavoring agent such aspeppermint, methyl salicylate or orange flavoring. When the dosage unitform is a capsule, it may contain, in addition to materials of the abovetype, a liquid carrier such as polyethylene glycol or a fatty oil. Otherdosage unit forms may contain other various materials which modify thephysical form of the dosage unit, for example, as coatings. Thus,tablets or pills may be coated with sugar, shellac, or other coatingagents. A syrup may contain, in addition to the present compounds,sucrose as a sweetening agent and certain preservatives, dyes andcolorings and flavors. Materials used in preparing these variouscompositions should be pharmaceutically pure and non-toxic in theamounts used.

For the purpose of oral and parenteral therapeutic administration, thecompounds f the present invention may be incorporated into a solution orsuspension. These preparations typically contain at least 0.1% of acompound of the invention, but may be varied to be between 0.1 and about90% of the weight thereof. The amount of the compound of Formula Ipresent in such compositions is such that a suitable dosage will beobtained. The solutions or suspensions may also include one or more ofthe following adjuvants: sterile diluents such as water for injection,saline solution, fixed oils, polyethylene glycols, glycerine, propyleneglycol or other synthetic solvents; antibacterial agents such as benzylalcohol or methyl paraben; antioxidants such as ascorbic acid or sodiumbisulfite; chelating agents such as ethylene diaminetetraacetic acid;buffers such as acetates, citrates or phosphates and agents for theadjustment of tonicity such as sodium chloride or dextrose. Theparenteral preparation can be enclosed in ampoules, disposable syringesor multiple dose vials made of glass or plastic. Preferred compositionsand preparations are able to be determined by one skilled in the art.

The compounds of the present invention may also be administeredtopically, and when done so the carrier may suitably comprise asolution, ointment, or gel base. The base, for example, may comprise oneor more of the following: petrolatum, lanolin, polyethylene glycols,bees wax, mineral oil, diluents such as water and alcohol, andemulsifiers, and stabilizers. Topical formulations may contain aconcentration of the Formula I or its pharmaceutical salt from about 0.1to about 10% w/v (weight per unit volume).

The compounds of Formula I are agonists of the M-1 muscarinic receptors.Moreover the compounds of Formula I are selective agonists of thatparticular muscarinic receptor. The compounds of the present inventionpossess particularly useful properties related to their bioavailability,pharmacokinetics, safety, and efficacy. Muscarinic agonists, includingtheir subtype binding profile, can be identified by the methods that arewell known in the art.

In one embodiment, the present invention provides methods of treatingdisorders associated with muscarinic receptors, comprising:administering to a patient in need thereof an effective amount of acompound of Formula I. Thus, the present invention contemplates thevarious disorders described to be treated herein and others which can betreated by such agonists as are appreciated by those skilled in the art.

A number of the disorders which can be treated by muscarinic agonistsare known according to established and accepted classifications, whileothers are not. For example, cognition is a complicated and sometimespoorly defined phenomenon. It is, however, widely recognized thatcognition includes various “domains.” These domains include short termmemory, long term memory, working memory, executive function, andattention.

It is understood that the compounds of the present invention are usefulfor treatment of disorders characterized by a deficit in any of thecognitive domains listed above or in other aspects of cognition. Thusthe term “cognitive disorders” is meant to encompass any disordercharacterized by a deficit in one or more cognitive domain, includingbut not limited to short term memory, long term memory, working memory,executive function, and attention.

One cognitive disorder to be treated by the present invention isage-related cognitive decline. This disorder is not well defined in theart, but includes decline in the cognitive domains, particularly thememory and attention domains, which accompany aging. Another cognitivedisorder is mild cognitive impairment. Again, this disorder is not welldefined in the art, but involves decline in the cognitive domains, andis believed to represent a group of patients the majority of which haveincipient Alzheimer's disease. Another cognitive disorder is cognitiveimpairment associated with schizophrenia. The relationship betweencognitive disturbances and other symptoms of schizophrenia is notclearly understood at present. It has been observed that some peopleexperience cognitive problems much before they develop positivesymptoms, while others acquire cognitive deterioration after the firstepisode and with subsequent relapses. Yet another cognitive disorder ischemotherapy-induced cognitive impairment. People who undergo cancerchemotherapy may experience a decline in cognitive function and thisdecline can be long lasting. Also, a wide variety of insults, includingstroke, ischemia, hypoxia, inflammation, infectious processes andcognitive deficits subsequent to cardiac bypass surgery and grafting,stroke, cerebral ischemia, spinal cord trauma, head trauma, perinatalhypoxia, fetal alcohol syndrome, cardiac arrest, and hypoglycemicneuronal damage, vascular dementia, multi-infarct dementia, amylotrophiclateral sclerosis, chemotherapy, and multiple sclerosis can result incognitive deficits as a sequella which can be treated according to thepresent invention.

Where the disorders which can be treated by muscarinic agonists areknown according to established and accepted classifications, theseclassifications can be found in various sources. For example, atpresent, the fourth edition of the Diagnostic and Statistical Manual ofMental Disorders (DSM-IV™) (1994, American Psychiatric Association,Washington, D.C.), provides a diagnostic tool for identifying many ofthe disorders described herein. Also, the International Classificationof Diseases, Tenth Revision (ICD-10), provides classifications for manyof the disorders described herein. The skilled artisan will recognizethat there are alternative nomenclatures, nosologies, and classificationsystems for disorders described herein, including those as described inthe DSM-IV and ICD-10, and that terminology and classification systemsevolve with medical scientific progress.

In particularly preferred embodiments, the present invention providesmethods of treating disorders selected from the group consisting of:cognitive disorders (including age-related cognitive disorder, mildcognitive impairment, cognitive impairment associated withschizophrenia, and chemotherapy-induced cognitive impairment), ADHD,mood disorders (including depression, mania, bipolar disorders),psychosis (in particular schizophrenia and schizophreniform disorder),dementia (including Alzheimer's disease, AIDS-induced dementia, vasculardementia, and dementia lacking distinctive histology), Parkinson'sdisease, Huntington's Chorea, pain (including acute pain and chronicpain), xerostomia (dry mouth), Lewy body disease (including diffuse Lewybody disease), aphasia (including primary aphasia and primary aphasiasyndromes), aphasia (including primary aphasia and primary aphasiasyndromes), hypotensive syndromes, and chronic colitis (includingCrohn's disease), comprising: administering to a patient in need thereofan effective amount of a compound of Formula I. That is, the presentinvention provides for the use of a compound of Formula I orpharmaceutical composition thereof for the treatment disordersassociated with muscarinic receptors.

It is recognized that the terms “treatment” and “treating” are intendedto include improvement of the symptomatology associated with each of thedisorders associated with muscarinic receptors described herein. Also,it is also recognized that one skilled in the art may affect thedisorders by treating a patient presently afflicted with the disordersor by prophylactic ally treating a patient believed to be susceptible tosuch disorders with an effective amount of the compound of Formula I.Thus, the terms “treatment” and “treating” are intended to refer to allprocesses wherein there may be a slowing, interrupting, arresting,controlling, or stopping of the progression of the disorders describedherein, but does not necessarily indicate a total elimination of allsymptoms, and is intended to include prophylactic treatment of suchdisorders.

It is understood that the present invention includes adjunctivetreatment of the disorders described herein. More specifically, thecompounds of Formula I are useful to treat disorders in which acognitive deficit is one of the symptoms in combination with a widevariety of other therapeutic agents, in particular, in combination withAMPA potentiators; with typical and atypical antipsychotics, includingolanzapine; with a variety of agents such as mGluR agonists, with NMDAantagonists, with IL 1-6 inhibitors, with other cholinergics, includingcholinesterase inhibitors, such as tacrine and donepezil, and compoundsthat inhibit amyloid protein processing, including inhibitors of amyloidprecursor protein processing and antibodies directed against amyloidproteins; with antidepressants, including SSRIs and SNRIs such asfluoxetine, paroxetine, and venlafaxine; and with anxiolytic agents;etc. It is believed that the combinations above are synergisticallybeneficial providing efficacy at doses that are a small fraction ofthose required to produce the same effect with the individualcomponents.

In accordance with the adjunctive treatments described above, thepresent invention also provides a product containing a compound ofFormula I and one or more therapeutic agents selected from the groupconsisting of AMPA potentiators; typical and atypical antipsychotics,including olanzapine; mGluR agonists; NMDA antagonists; IL 1-6inhibitors; cholinesterase inhibitors, such as tacrine and donepezil;compounds that inhibit amyloid protein processing, including inhibitorsof amyloid precursor protein processing and antibodies directed againstamyloid proteins; antidepressants, including SSRIs and SNRIs such asfluoxetine, paroxetine, and venlafaxine; and anxiolytic agents as acombined preparation for simultaneous, separate or sequentialadministration in the treatment of disorders in which a cognitivedeficit is one of the symptoms. In another embodiment the presentinvention also provides for the use of a compound of Formula I togetherwith one or more therapeutic agents selected from AMPA potentiators;typical and atypical antipsychotics, including olanzapine; mGluRagonists; NMDA antagonists; IL 1-6 inhibitors; cholinesteraseinhibitors, such as tacrine and donepezil; compounds that inhibitamyloid protein processing, including inhibitors of amyloid precursorprotein processing and antibodies directed against amyloid proteins;antidepressants, including SSRIs and SNRIs such as fluoxetine,paroxetine, and venlafaxine; and anxiolytic agents for the manufactureof a medicament as a combined preparation for simultaneous, separate orsequential administration in the treatment of disorders in which acognitive deficit is one of the symptoms.

As used herein, the term “simultaneous, separate or sequentialadministration” means that the two or more therapeutic agents areadministered within a time frame which ensures that all of thetherapeutic agents will provide some therapeutic activity at aparticular point in time. That is to say, the therapeutic activitiesshould at least overlap to some degree although they need not becoterminus.

As used herein, the term “patient” includes a mammal which is afflictedwith one or more disorders associated with muscarinic receptors. It isunderstood that guinea pigs, dogs, cats, rats, mice, horses, cattle,sheep, pigs, and humans are examples of animals within the scope of themeaning of the term.

As used herein, the term “effective amount” of a compound of Formula Irefers to an amount, that is, the dosage which is effective in treatingthe disorders described herein.

An effective amount can be readily determined by the attendingdiagnostician, as one skilled in the art, by the use of conventionaltechniques and by observing results obtained under analogouscircumstances. In determining an effective amount, the dose of acompound of Formula I, a number of factors are considered by theattending diagnostician, including, but not limited to: the compound ofFormula I to be administered; the co-administration of other therapies,if used; the species of mammal; its size, age, and general health; thespecific disorder involved; the degree of involvement or the severity ofthe disorder; the response of the individual patient; the mode ofadministration; the bioavailability characteristics of the preparationadministered; the dose regimen selected; the use of other concomitantmedication; and other relevant circumstances.

An effective amount of a compound of Formula I is expected to vary fromabout 0.01 milligram per kilogram of body weight per day (mg/kg/day) toabout 50 mg/kg/day, and preferable from 0.1 milligram per kilogram ofbody weight per day (mg/kg/day) to about 20 mg/kg/day. More preferredamounts can be determined by one skilled in the art.

Of the disorders to be treated according to the present invention anumber are particularly preferred. Particularly preferred disordersinclude the treatment of cognitive disorders (particularly mildcognitive impairment and cognitive impairment associated withschizophrenia), Alzheimer's disease, and psychosis, includingschizophrenia.

A number of preclinical laboratory animal models have been described forthe disorders described herein.

EXAMPLE A

Radial Arm Maze

The delayed non-match to sample task has been used to study the effectof drugs on memory retention (Pussinen, R. and Sirvio, J. J ofPsychopharm 13: 171-179(1999); Staubli, U., et al. Proc Natl Acad Sci91: 777-781(1994)) in the eight arm radial maze.

Well-trained rats were allowed to retrieve food rewards from fourrandomLy selected arms of the maze (sampling phase). Some time later,the rats were exposed to eight open arms and were tested for theirability to remember and avoid the arms they had previously entered toobtain food. Re-entry into an arm that was baited during the samplingsession was counted as a reference error, whereas entry into the samearm more than once during the retention session was counted as workingerror. The total (reference+working) number of errors made during theretention test increases with increasing delay periods. For example,young male rats made 0.66 (+0.4) errors at a 1 minute delay, 2 (+0.5)errors at a one hour delay, and 3.95 (+0.2) errors at a seven hour delay(observations of this lab).

Male Sprague-Dawley rats were individually housed and maintained on a 12h light-dark cycle (lights on at 6 am). The rats were given free accessto water and maintained at 85% of their free-feeding weight bysupplemental feedings of Purina Lab Chow.

The rats were initially trained to search for food at the end of each ofthe eight arms. Once the rats had reached the criteria of no more thantwo errors (i.e. entering the same arm more than once during a session)on three consecutive days, a delay of one minute was imposed between thefourth and the fifth arm choices. This training ensured that the ratswere thoroughly familiar with the procedural aspects of the task beforeany drugs were administered. Once stable performance had been obtainedon the delay task (i.e. no more than one error was made on threeconsecutive days), drug and vehicle tests commenced using a seven hourdelay period. A novel set of arms was baited each day for each rat andthe maze was thoroughly cleaned during the delay period.

During the sampling session, each rat was placed on the center platformwith access to all eight arms of the maze blocked. Four of the eightarms were randomly selected and baited with food. The gates of thebaited arms were raised and the rat was allowed five minutes to obtainthe food at the end of each of the four arms. As soon as the rat hadobtained the food, it was removed, administered vehicle or various dosesof compounds, and placed back in its home cage. Seven hours later(retention session), the rat was placed back onto the center platformwith access to all eight arms blocked. The four arms that werepreviously baited during the sampling session, were baited and the gatesto all eight arms were raised. The rat was allowed five minutes toobtain the remaining four pieces of food. A n entry into a non-baitedarm or a re-entry into a previously visited arm was counted as an error.Significance (p<0.05) was determined using a repeated measure ANOVAfollowed by a Dunnett's test for comparison with control.

In order to compare test compounds with standards, scopolamine andtacrine were administered s.c. immediately after the sampling phase. Theeffects of scopolamine, a known amnesic, were tested after a three-hourdelay, whereas the effect of tacrine, a cholinesterase inhibitor used inthe treatment of Alzheimer's disease was tested after a six-hour delay.Scopolamine disrupted retention after a three-hour delay in adose-related fashion. Tacrine significantly improved retention after asix-hour delay at 10, but not at 3 mg/kg.

EXAMPLE B

Acquisition in the Radial Maze 8-Arm Radial Maze Acquisition

A prominent early feature of Alzheimer's disease (AD) symptomology is apronounced deficit in declarative memory (R. W. Parks, R. F. Zec & R. S.Wilson (Eds.), Neuropsychology of Alzheimer's disease and otherdementias. NY: Oxford University Press pp. 3-80 (1993).

As the disease progresses, other domains of cognition become severelyaffected as well. Among the brain regions affected early in theprogression of Alzheimer's disease is the hippocampus, which is acritical neural substrate for declarative memory. Differences in thepattern of hippocampal neuronal loss in normal aging and Alzheimer'sdisease. Lancet, 344: 769-772(1994). One behavioral test that is oftenused to assess hippocampal function in animal models is the 8-arm radialmaze (Olton D. S. The radial arm maze as a tool in behavioralpharmacology. Physiology & Behavior, 40: 793-797 (1986)).

Lesions or pharmacological blockade of the hippocampus disruptperformance of this task. Moreover, aged animals generally show deficitsin this task (Porsolt R. D., Roux S. & Wettstein J. G. Animal models ofdementia. Drug Development Research, 35: 214-229(1995)).

In this test of spatial learning and memory, a hungry rat is placed inthe center of the maze and allowed to traverse the maze in search offood located at the end of each runway arm. In this version of the maze,the rat learns a win-shift strategy in which a visited arm is notreplaced. Therefore, the most efficient foraging strategy is to visiteach arm once. The version of the maze also taps into general learningprocesses as the rat is naive to the maze on day one of the four dayexperiment.

Upon arrival, male Sprague Dawley®, rats were individually housed in aregular light-cycle colony room and allowed to acclimate for at least 4days prior to testing. Each rat was reduced to and maintained at 85% oftheir target body weight throughout the experiment. Proper body weightwas maintained by adjusting the allotment of lab chow based on acombination of age and the rat's daily bodyweight reading.

A session began with an individual rat being placed into the hub of themaze and then all guillotine doors were raised, allowing free access toall areas of the maze. A food hopper was located at the end of each ofthe 8 runway arms and a single food pellet was placed in each foodhopper. Each daily session terminated when either all 8 food-hoppers hadbeen visited or when the rat timed out (15 minutes on Day 1: 5 minuteson Days 2-4). The number of arm entries was recorded. Errors werecounted as repeat arm entries or failures to visit an arm in the sessionperiod. An animal was excluded from the study if it failed to visit atleast one arm on Day 1, 2 arms on Day 2, and at least 4 arms on Days 3 &4.

Each rat was pseudo-randomly assigned to either a vehicle or drug groupand received the same treatment throughout the experimental period.Vehicle consisted of 5% acacia within sterile water. Injections wereadministered subcutaneously 20-30 minutes prior to each daily session.

In this acquisition task, vehicle-treated animals do not consistentlyshow significant acquisition of maze learning as compared to the numberof errors committed on Day 1. We have found that in compounds thatfacilitate acquisition of maze learning, the effects are often notobserved until the fourth day of training. Therefore, results consistedof total Day 4 errors across treatment groups.

EXAMPLE C

Functional Mobilization of Intracellular Calcium

CHO cells expressing muscarinic subtypes (M1-M5) are grown as monolayersin DMEM:F-12 (3:1), 10% FBSnz, 20 mM HEPES, 1% pen/strep, 250 μg/mL G418(GibcoBRL #10131-027). Cells are maintained under 95%/5% O₂/CO₂ andpassaged every 3-4 days. Cells are plated 24 hours in advance of theassay at a density of 50,000/well and 48 hours in advance at a densityof 25,000/well (100 μL/well) in Costar black-walled, clear-bottomed 96well plates (Costar #3603). Cells are then incubated with minimumessential medium containing the cytoplasmic Ca²⁺ indicator, Fluo-3 (1 mMFluo mixed 1:1 with 20% pluronic acid, then diluted to 5 μM finalconcentration in growth and supplemented with 2.5 mM, 50 μL/well) at 37°C. in an environment containing 5% CO₂ for 60 minutes. Cells are washedtwice with 100 μL/well of wash buffer containing Hanks Balanced SaltSolution (HBSS) without phenol red (1×) (GibcoBRL #14065-056), 20 mMHEPES (Sigma #P8761), and Probenecid (2.5 mM) (100×: 1:100). For theassay, 100 μL is added to each well (100 μL of 2× drug will be added bythe FLIPR). Plates are washed three times using a LabSystems multidropand residual buffer is removed. Plates are also blotted on paper towelsto remove remaining compound.

Compounds are prepared 2× (100 μL of drug added to 100 μL of assaybuffer present in the well) in assay buffer containing 2% DMSO, HBSSwithout phenol red (1×) (GibcoBRL #14065-056), 20 mM HEPES (Sigma#P8761), and Probenecid (2.5 mM) (100×: 1:100).

The plates were then placed into a FLIPR instrument (fluorometricimaging plate reader system, Molecular Devices, Sunnyvale, Calif.) tomonitor cell fluorescence (λ_(EX)═488 nm, λ_(EM)═540 nm) before andafter the addition of compounds.

The selectivity of the M1 agonists are evaluated by screening across theother muscarinic receptor subtypes (M2, M3, M4 and M5) in a similarmanner. Compounds are also screened across a number of protein targetsas well as the structurally related G protein-coupled receptor (GPCR)targets to insure selectivity for the M1 receptor.

EXAMPLE D

Functional GTP Binding

Cell Culture: CHO cells transfected with human M1-M5 receptors weregrown either in suspension or in monolayer. For suspension culturescells were grown in roller bottles with constant agitation at 37° C. and5% CO₂ using Dulbecco's modified Eagles medium/F-12 (3:1) culture mediumsupplemented with 5% fetal bovine serum, 50 μg/mL tobramycin, and 20 mMHEPES. Monolayer cultures were grown in T-225 flasks at 37° C. and 5%CO2 in Dulbecco's modified Eagles medium supplemented with 10% fetalbovine serum and 100,000 U/liter of penicillin/streptomycin. Cells wereharvested using trypsin-free dissociation media at 95% confluence andwere collected by centrifugation and stored at 80° C. Cells stablyexpressing human muscarinic receptors were obtained from the NationalInstitutes of Health.

Membrane Preparation: Cell pellets were thawed and resuspended in 20volumes of 20 mM sodium phosphate buffer, pH 7.4, and were homogenizedtwice for 30 seconds at high speed using a Tissuemizer. Homogenates werecentrifuged at 200 g for 15 minutes at 4° C. The supernatant was removedand reserved on ice. This procedure was repeated twice and the pooledsupernatants were then centrifuged at 40,000 g for 45 minutes at 4° C.Membranes were suspended at 5 mg protein/mL and were stored at 80° C.Unless indicated otherwise in the figure legends, membranes from M1, M2,and M4 cells were prepared from cells grown in suspension, whereas thosefrom M3 and M5 cells were from cells grown in monolayer. Receptordensities (pmol mg1 membrane protein) were 9.3, 0.7, 0.6, 0.9, and 4.8for M1-M5 receptors, respectively.

Striatal tissue from male Sprague-Dawley rats was homogenized by hand in10 volumes of 10 mM HEPES and 1 mM EGTA, pH 7.4, containing Completeprotease inhibitor cocktail, 1 mM dithiothreitol, and 10% sucrose. Thehomogenate was diluted 6-fold and centrifuged at 1000 g for 10 minutesat 4° C. The supernatant was saved and the pellet rehomogenized andcentrifuged as above. The combined supernatants were centrifuged at11,000 g for 20 minutes. The resulting pellet was homogenized in 40volumes of 10 mM HEPES and 1 mM EGTA, pH 7.4, containing 1 mMdithiothreitol and 1 mM MgCl₂, and was centrifuged at 27,000 g for 20minutes. The resulting pellet was suspended in the same buffer at aprotein concentration of 1.5 mg/mL and aliquots were frozen and storedat 80° C.

GTPγ³⁵S Binding: Assays were run in 20 mM HEPES, 100 mM NaCl, and 5 mMMgCl₂ at pH 7.4 in a final volume of 200 μl in 96-well Costar plates at25° C. One hundred microliters of membrane preparation (25 μg proteinper well for cell membranes and 9-15 μg per well for brain membranes)containing the appropriate concentration of GDP was added followed byaddition of 50 μl of buffer±agonists and antagonists being testedfollowed by 50 μl of GTPγ³⁵S to provide a final concentration in theassay of 200 pM for CHO membranes and 500 pM for brain membranes. ForCHO membranes, 0.1 μM GDP was used for M1, M3, and M5 receptor assays,whereas 1 μM GDP was used for M2 and M4 assays. For brain membranes, 0.1μM GDP was used in assays carried out with anti-Gαq/11, whereas 50 μMGDP was used for assays using anti-Gαi(1-3) and anti-Gαo. CHO cellmembranes were incubated for 30 minutes at 25° C. with agonists andantagonists followed by addition of GTPγ³⁵S and incubation for anadditional 30 minutes. Brain membranes were incubated for 20 minutes at25° C. with agonists and antagonists followed by addition of GTPγ³⁵S andincubation for an additional 60 minutes. Preincubation was employed toensure that agonists and antagonists were at equilibrium during thelabeling period.

To determine total membrane binding, 50 μL of suspended wheat germagglutinin (WGA)-coated SPA beads was added. After 15 minutes, plateswere centrifuged at 1000 g for 15 minutes and radioactivity wasdetermined using a Wallac plate counter. For determining binding tospecific G proteins, ³⁵S-labeled membranes were solubilized for 30minutes with 0.27% Nonidet P-40 (20 μL/well of a solution containing 1.5mL of 10% Nonidet P-40 for every 3.5 mL assay buffer) followed byaddition of desired antibody (10 μl/well) to provide a final dilution of1/400 to 1/100 and incubation for an additional 60 minutes. Fiftymicroliters of suspended anti-IgG-coated SPA beads was added per well,plates were incubated for 3 hours, and then were centrifuged andradioactivity determined as above. Each bottle of WGA-coated SPA beadswas suspended in 10 mL of assay buffer and each bottle ofanti-IgG-coated SPA beads was suspended in 20 mL of assay buffer.Protein was determined using the bicinchoninic acid assay.

Materials: ³⁵S-GTPγS (1000-1200 Ci/mmol), anti-rabbit-IgG andanti-mouse-IgG-coated SPA beads, and WGA-coated SPA beads were obtainedfrom Amersham (Arlington Heights, Ill.). Rabbit anti-Gαq/11 and rabbitanti-Gαi(1-3) were from Santa Cruz Biotechnologies (Santa Cruz, Calif.).Mouse monoclonal anti-Gαo was from Chemicon (Temecula, Calif.).Oxotremorine M and pirenzepine were from Research Biochemicals Inc.(Natick, Mass.).11-{[2-((Diethylamino)methyl)-1-piperidinyl]acetyl}-5,11-dihydro-6H-pyrido[2,3b][1,4]benzodiazepin-6-one(AFDX 116) was synthesized at Eli Lilly. Complete protease inhibitorcocktail and 10% Nonidet P-40 were from Boehringer Mannheim(Indianapolis, Ind.).

The selectivity of the M1 agonists are evaluated by screening across theother muscarinic receptor subtypes (M2, M3, M4 and M5). Compounds arealso screened across a number of protein targets as well as thestructurally related G protein-coupled receptor (GPCR) targets to insureselectivity for the M1 receptor.

1. A compound of the Formula

wherein Q, X, Y, and Z are independently selected from the groupconsisting of CR¹ and N, provided that no more than two of Q, X, Y, andZ are N and at least two of Q, X, Y, and Z are CH; or Y is CH, Z is CH,and the moiety “Q═X” represents “S” to form a thiophene ring; R¹ isindependently at each occurrence selected from the group consisting ofhydrogen, halogen, C₁-C₄ alkoxy, and C₁-C₄ alkyl; R² is selected fromthe group consisting of halogen; C₁-C₄ alkoxy; C₁-C₄ alkyl; C₃-C₈cycloalkyl; cyano; trifluoromethyl; pyridinyl optionally substitutedwith one to two substituents independently selected from the groupconsisting of halogen, C₁-C₄ alkoxy, and C₁-C₄ alkyl; thienyl optionallysubstituted with one substituent selected from the group consisting ofhalogen, C₁-C₄ alkoxy, and C₁-C₄ alkyl; phenyl optionally substitutedwith from one to three substituents independently selected from thegroup consisting of halogen, C₁-C₄ alkoxy, C₁-C₄ alkyl, trifluoromethyl,and cyano; and pyrrolyl optionally substituted with one to twosubstituents independently selected from the group consisting ofhalogen, C₁-C₄ alkoxy, and C₁-C₄ alkyl; R³ is a radical of the formula(Z)-(Y)—(X)— wherein X is selected from the group consisting of

and a straight-chain C₁-C₄ alkandiyl optionally substituted with methyl,geminal dimethyl, or phenyl; Y is selected from the group consisting ofO and S; and Z is selected from the group consisting of C₁-C₆ alkyl;C₃-C₈ cycloalkyl optionally substituted with one to three substituentsindependently selected from the group consisting of halogen, C₁-C₄alkoxy, C 1-C₄ alkyl, trifluoromethyl, cyano, and nitro; phenyloptionally substituted with one to three substituents independentlyselected from the group consisting of halogen, C₁-C₄ alkoxy, C₁-C₄alkyl, trifluoromethyl, cyano, and nitro; naphthyl optionallysubstituted with one to three substituents independently selected fromthe group consisting of halogen, C₁-C₄ alkoxy, C₁-C₄ alkyl,trifluoromethyl, cyano, and nitro; heteroaryl optionally substitutedwith one or two substituents independently selected from the groupconsisting of halogen, C₁-C₄ alkoxy, and C₁-C₄ alkyl; and heterocycleoptionally substituted with one or two substituents independentlyselected from the group consisting of halogen, C₁-C₄ alkoxy, and C₁-C₄alkyl; R^(a) is selected from the group consisting of hydrogen andmethyl; R⁴ is selected from the group consisting of hydrogen, hydroxy,and fluoro; R⁵ is selected from the group consisting of hydrogen,halogen, C₁-C₄ alkoxy, and C₁-C₄ alkyl; R^(b) is selected from the groupconsisting of hydrogen, methyl, and ethyl; and m is one or two; orpharmaceutically acceptable addition salts thereof.
 2. A compound ofclaim 1 wherein R⁵ is hydrogen, R⁴ is hydroxy, m is one, and which hasthe trans stereochemistry at the 1- and 2-position shown below:


3. A compound according to claim 1 wherein Q, X, Y, and Z are each CH.4. A compound according to claim 2 wherein Q, X, Y, and Z are each CH.5. A compound according to claim 1 wherein one of Q, X, Y, and Z is CFand the others are CH.
 6. A compound according to claim 2 wherein one ofQ, X, Y, and Z is CF and the others are CH.
 7. A compound according toclaim 1 wherein Q is CF, and X, Y, and Z are each CH.
 8. A compoundaccording to claim 1 wherein R² is phenyl optionally substituted withfrom one to three substituents independently selected from the groupconsisting of halogen, C₁-C₄ alkoxy, C₁-C₄ alkyl, trifluoromethyl, andcyano.
 9. A compound according to claim 1 wherein R² is phenyl.
 10. Apharmaceutical composition comprising a compound of claim 1 and apharmaceutically acceptable diluent.
 11. A method of treatingAlzheimer's disease, comprising: administering to a patient in needthereof an effective amount of a compound of claim
 1. 12. A method oftreating schizophrenia, comprising: administering to a patient in needthereof an effective amount of a compound of claim 1.